Publications

isolation of a peptide inhibitor of human rhinovirus.	cell culture-based transdominant genetic techniques provide new methods for discovering peptide/rna modulators of cellular pathways. we applied this technology to isolate a peptide inhibitor of human rhinovirus. a green fluorescent protein (gfp)-scaffolded library of cdna fragments was expressed in hela cells from a retroviral vector and screened for inhibitors of rhinovirus-mediated cell killing. a dna clone, i421, increased cell survival in an hrv14 challenge assay from less than 0.5% to great ...	2003	12951031
demonstrating the intrinsic ion channel activity of virally encoded proteins.	this review summarizes the types of evidence that can be invoked in order to demonstrate that a virally encoded protein possesses ion channel activity that is intrinsic to the life cycle of the virus. ion channel activity has been proposed to be a key step in the life cycle of influenza virus, and the protein responsible for this activity has been proposed to be the m2 protein encoded by the virus. this review contrasts the evidence supporting the conclusion that the a/m2 protein of influenza a ...	2003	12972153
imidazo[1,2-b]pyridazines, novel nucleus with potent and broad spectrum activity against human picornaviruses: design, synthesis, and biological evaluation.	a novel structural class of picornavirus inhibitors comprising an imidazo[1,2-b]pyridazine nucleus was discovered. 2-aminoimidazo[1,2-b]pyridazines (6d, (e/z)-7b, (e)-7d, (z)-7d, (e/z)-8b, (e)-10b, (e)-13a, (z)-13a, (e)-13b, (z)-13b, (e)-13c, and (z)-13c) were designed and synthesized in an effort to identify potent broad spectrum antirhinoviral agents. a practical synthetic route to this chemical scaffold has been developed. the target compounds were evaluated in a plaque reduction assay and in ...	2003	13678411
mouse respiratory epithelial cells support efficient replication of human rhinovirus.	human rhinoviruses (hrv) are responsible for the majority of virus infections of the upper respiratory tract. furthermore, hrv infection is associated with acute exacerbation of asthma and other chronic respiratory diseases of the lower respiratory tract. a small animal model of hrv-induced disease is required for the development of new therapies. however, existing mouse models of hrv infection are difficult to work with and until recently mouse cell lines were thought to be generally non-permis ...	2003	13679617
synthesis and evaluation of tripeptidyl alpha-ketoamides as human rhinovirus 3c protease inhibitors.	we describe herein the synthesis and biological evaluation of a series of tripeptidyl alpha-ketoamides as human rhinovirus (hrv) 3c protease inhibitors. the most potent inhibitor discussed in this manuscript, 4i, exhibited impressive enzyme inhibitory activity as well as antiviral activity against hrv-14.	2003	14505664
human rhinovirus capsid dynamics is controlled by canyon flexibility.	quantitative enzyme accessibility experiments using nano liquid chromatography electrospray mass spectrometry combined with limited proteolysis and isotope-labeling was used to examine the dynamic nature of the human rhinovirus (hrv) capsid in the presence of three antiviral compounds, a neutralizing fab, and drug binding cavity mutations. using these methods, it was found that the antivirals win 52084 and picovir (pleconaril) stabilized the capsid, while dansylaziridine caused destabilization. ...	2003	14517058
functional conservation of the hydrophobic domain of polypeptide 3ab between human rhinovirus and poliovirus.	in this study we exchanged portions of the poliovirus type 1 (pv1) hydrophobic domain within the membrane-associated polypeptide 3ab for the analogous sequences from human rhinovirus 14 (hrv14). the sequence exchanges were based upon a previous report in which the 22 amino acid hydrophobic region was subdivided into two domains, i and ii, the latter of which was shown to be required for membrane association (j. biol. chem. 271 (1996), 26810). using these divisions, the hrv14 sequences were clone ...	2003	14517095
structure-based design, synthesis, and biological evaluation of irreversible human rhinovirus 3c protease inhibitors. 8. pharmacological optimization of orally bioavailable 2-pyridone-containing peptidomimetics.	the optimization of the pharmacokinetic performance of various 2-pyridone-containing human rhinovirus (hrv) 3c protease (3cp) inhibitors following oral administration to either beagle dogs or cm-monkeys is described. the molecules described in this work are composed of a 2-pyridone-containing peptidomimetic binding determinant and an alpha,beta-unsaturated ester michael acceptor moiety which forms an irreversible covalent adduct with the active site cysteine residue of the 3c enzyme. modificatio ...	2003	14521419
rna cleaving '10-23' dnazymes with enhanced stability and activity.	'10-23' dnazymes can be used to cleave any target rna in a sequence-specific manner. for applications in vivo, they have to be stabilised against nucleolytic attack by the introduction of modified nucleotides without obstructing cleavage activity. in this study, we optimise the design of a dnazyme targeting the 5'-non-translated region of the human rhinovirus 14, a common cold virus, with regard to its kinetic properties and its stability against nucleases. we compare a large number of dnazymes ...	2003	14530446
phase ii, randomized, double-blind, placebo-controlled studies of ruprintrivir nasal spray 2-percent suspension for prevention and treatment of experimentally induced rhinovirus colds in healthy volunteers.	human rhinovirus (hrv) infections are usually self-limited but may be associated with serious consequences, particularly in those with asthma and chronic respiratory disease. effective antiviral agents are needed for preventing and treating hrv illnesses. ruprintrivir (agouron pharmaceuticals, inc., san diego, calif.) selectively inhibits hrv 3c protease and shows potent, broad-spectrum anti-hrv activity in vitro. we conducted three double-blind, placebo-controlled clinical trials in 202 healthy ...	2003	14638501
activity of a type 1 picornavirus internal ribosomal entry site is determined by sequences within the 3' nontranslated region.	we have proposed a cancer treatment modality based on poliovirus chimeras replicating under the translational control of an internal ribosomal entry site (ires) derived from human rhinovirus type 2. insertion of the heterologous ires causes a neuron-specific propagation deficit and eliminates neurovirulence inherent in poliovirus without affecting viral growth in cells derived from malignant gliomas. we now report the elucidation of a molecular mechanism responsible for the cell type-specific de ...	2003	14645707
the human rhinovirus internal cis-acting replication element (cre) exhibits disparate properties among serotypes.	it has been reported previously that the human rhinovirus 14 (hrv-14) rna genome contains a cis-acting replication element (cre) that maps to the capsid coding (p1) sequence [19]. further characterization of the hrv-14 cre in the present study established that by moving the cre stem-loop structure downstream, adjacent to the 3'ncr, that its position is not critical for function. when the p1 sequences of two closely related serotypes of hrv-14 were analyzed for the presence of a cre, both hrv-3 a ...	2003	14648294
biochemical and genetic studies of the vpg uridylylation reaction catalyzed by the rna polymerase of poliovirus.	the first step in poliovirus (pv) rna synthesis is the covalent linkage of ump to the terminal protein vpg. this reaction can be studied in vitro with two different assays. the simpler assay is based on a poly(a) template and requires synthetic vpg, purified rna polymerase 3d(pol), utp, and a divalent cation. the other assay uses specific viral sequences [cre(2c)] as a template for vpg uridylylation and requires the addition of proteinase 3cd(pro). using one or both of these assays, we analyzed ...	2003	12502805
virus-ligand interactions: identification and characterization of ligand binding by nmr spectroscopy.	we demonstrate the detection and characterization of ligand binding to viruses via nmr. to illustrate the methodology, the interaction of an antiviral compound with human rhinovirus serotype 2 (hrv2) was investigated. specific interaction of a capsid-binding inhibitor and native hrv2 was monitored utilizing saturation transfer difference (std) nmr. std nmr experiments at atomic resolution allowed those regions of the ligand that are involved in the interaction with the virus to be determined. th ...	2003	12515488
human rhinovirus selectively modulates membranous and soluble forms of its intercellular adhesion molecule-1 (icam-1) receptor to promote epithelial cell infectivity.	human rhinoviruses are responsible for many upper respiratory tract infections. 90% of rhinoviruses utilize intercellular adhesion molecule-1 (icam-1) as their cellular receptor, which also plays a critical role in recruitment of immune effector cells. two forms of this receptor exist; membrane-bound (micam-1) and soluble icam-1 (sicam-1). the soluble receptor may be produced independently from the membrane-bound form or it may be the product of proteolytic cleavage of micam-1. the ratio of airw ...	2003	12551926
structure and function analysis of the poliovirus cis-acting replication element (cre).	the poliovirus cis-acting replication element (cre) templates the uridylylation of vpg, the protein primer for genome replication. the cre is a highly conserved structural rna element in the enteroviruses and located within the polyprotein-coding region of the genome. we have determined the native structure of the cre, defined the regions of the structure critical for activity, and investigated the influence of genomic location on function. our results demonstrate that a 14-nucleotide unpaired t ...	2003	12554882
unr is required in vivo for efficient initiation of translation from the internal ribosome entry sites of both rhinovirus and poliovirus.	translation of picornavirus rnas is mediated by internal ribosomal entry site (ires) elements and requires both standard eukaryotic translation initiation factors (eifs) and ires-specific cellular trans-acting factors (itafs). unr, a cytoplasmic rna-binding protein that contains five cold-shock domains and is encoded by the gene upstream of n-ras, stimulates translation directed by the human rhinovirus (hrv) ires in vitro. to examine the role of unr in translation of picornavirus rnas in vivo, w ...	2003	12610110
viral receptor blockage by multivalent recombinant antibody fusion proteins: inhibiting human rhinovirus (hrv) infection with cfy196.	viral receptor blockage by monoclonal antibodies (mabs) is a common strategy to inhibit viral infection; however, no drug candidate acting by this mechanism has reached the market so far. analysis of the experimental and clinical data on a receptor-blocking mab, mab 1a6, against human rhinovirus (hrv) major receptor intercellular adhesion molecule 1 (icam-1) reveals that insufficient avidity of the mab is probably the key factor to blame for its unsatisfactory clinical efficacy. in this article, ...	2004	14657090
small interfering rna molecules as potential anti-human rhinovirus agents: in vitro potency, specificity, and mechanism.	rna silencing or interference (rnai) is a sequence-specific, post-transcriptional process of mrna degradation. the degradation of target gene mrna can be induced by short dsrna molecules (21-25-nt) corresponding to the sequence of the target gene to be silenced. short dsrna molecules have been shown to be very effective in inducing rna silencing in several human cell lines. in this study, we have shown that short dsrna molecules corresponding to the human rhinovirus-16 (hrv-16) genome induce eff ...	2004	14670593
genetic evidence for an interaction between a picornaviral cis-acting rna replication element and 3cd protein.	internally located, cis-acting rna replication elements, termed cres, are essential for replication of the genomes of picornaviruses such as human rhinovirus 14 (hrv-14) and poliovirus because they template uridylylation of the protein primer, vpg, by the polymerase 3d(pol). these cres form stem-loop structures sharing a common loop motif, and the hrv-14 cre can substitute functionally for the poliovirus cre in both uridylylation in vitro and rna replication in vivo. we show, however, that the p ...	2004	14711816
binding of fluorescent dye to genomic rna inside intact human rhinovirus after viral capsid penetration investigated by capillary electrophoresis.	ribogreen is used for concentration measurements of rna. upon binding to the rna, an approximately 1000-fold increase in sensitivity in comparison with the uv absorbance of the free polynucleotide is observed. in the present work, we demonstrate that this dye can penetrate in a time- and temperature-dependent manner the intact viral capsids of human rhinovirus serotypes 2 and 14, where it forms a fluorescent complex with the viral rna. capillary electrophoresis with laser-induced fluorescence de ...	2004	14961716
human rhinovirus type 2-antibody complexes enter and infect cells via fc-gamma receptor iib1.	hela cells were stably transfected with a cdna clone encoding the b1 isoform of the mouse fcgammarii receptor (hereafter referred to as hela-fcrii cells). the receptor was expressed at high level at the plasma membrane in about 90% of the cells. these cells bound and internalized mouse monoclonal virus-neutralizing antibodies 8f5 and 3b10 of the subtype immunoglobulin g2a (igg2a) and igg1, respectively. binding of the minor-group human rhinovirus type 2 (hrv2) to its natural receptors, members o ...	2004	14990693
cryoelectron microscopy analysis of the structural changes associated with human rhinovirus type 14 uncoating.	release of the human rhinovirus (hrv) genome into the cytoplasm of the cell involves a concerted structural modification of the viral capsid. the intracellular adhesion molecule 1 (icam-1) cellular receptor of the major-group hrvs and the low-density lipoprotein (ldl) receptor of the minor-group hrvs have different nonoverlapping binding sites. while icam-1 binding catalyzes uncoating, ldl receptor binding does not. uncoating of minor-group hrvs is initiated by the low ph of late endosomes. we h ...	2004	14990711
numa and nuclear lamins are cleaved during viral infection--inhibition of caspase activity prevents cleavage and rescues hela cells from measles virus-induced but not from rhinovirus 1b-induced cell death.	nuclear matrix is a structural framework of important nuclear processes. we studied the effect of two different types of viral infections on nuclear matrix. hela cells were infected with human rhinovirus 1b (hrv 1b) or measles virus (mv), and nuclear mitotic apparatus protein (numa) and lamins a/c and b were used as markers for internal nuclear matrix and peripheral nuclear lamina, respectively. we show that numa, lamins, and poly(adp-ribose) polymerase-1 are cleaved during viral infection in a ...	2004	15003865
monitoring rna release from human rhinovirus by dynamic force microscopy.	human rhinoviruses were imaged under physiological conditions by dynamic force microscopy. topographical images revealed various polygonal areas on the surfaces of the 30-nm viral particles. rna release was initiated by exposure to a low-ph buffer. the lengths of the rnas that were released but still connected to the virus capsid varied between 40 and 330 nm, whereas rna molecules that were completely released from the virus were observed with lengths up to 1 micro m. fork-like structure element ...	2004	15016841
vp1 sequencing of all human rhinovirus serotypes: insights into genus phylogeny and susceptibility to antiviral capsid-binding compounds.	rhinoviruses are the most common infectious agents of humans. they are the principal etiologic agents of afebrile viral upper-respiratory-tract infections (the common cold). human rhinoviruses (hrvs) comprise a genus within the family picornaviridae. there are >100 serotypically distinct members of this genus. in order to better understand their phylogenetic relationship, the nucleotide sequence for the major surface protein of the virus capsid, vp1, was determined for all known hrv serotypes an ...	2004	15016887
human rhinovirus infection induces airway epithelial cell production of human beta-defensin 2 both in vitro and in vivo.	we hypothesized that airway epithelial cells, the primary site of human rhinovirus (hrv) infection, provide a link between the innate and specific immune response to hrv via production of human beta-defensin (hbd)-2, a potent in vitro attractant and activator of immature dendritic cells. infection of primary cultures of human epithelial cells with several hrv serotypes induced expression of hbd-2 mrna and protein, indicating that hbd-2 production was independent of viral receptor usage or mechan ...	2004	15034083
synthesis and antiviral activity of novel fluorinated 2',3'-dideoxynucleosides.	a series of 5-(trifluoroethoxymethyl)-2',3'-dideoxyuridines and 5-[bis(trifluoroethoxy)-methyl]-2',3'-dideoxyuridines have been prepared and screened for antiviral activity. the conformations of these compounds are discussed on the bases of noe studies and the mo calculations. modelling and noe studies suggest both syn- and anti conformations for these 5-(2,2,2-trifluoroethoxymethyl)- and 5-[bis(2,2,2-trifluoroethoxy)-methyl]- derivatives. the noe parameters are also suggested to be more attribu ...	2004	15043133
x-ray structure of a minor group human rhinovirus bound to a fragment of its cellular receptor protein.	although many viral receptors have been identified, the ways in which they interact with their cognate viruses are not understood at the molecular level. we have determined the x-ray structure of a complex between calcium-containing modules of the very low-density lipoprotein receptor and the minor group human rhinovirus hrv2. the receptor binds close to the icosahedral five-fold vertex, with only one module per virus protomer. the binding face of this module is defined by acidic calcium-chelati ...	2004	15064754
role of nasal nitric oxide in the resolution of experimental rhinovirus infection.	human rhinovirus (hrv) infections are associated with exacerbations of asthma, chronic obstructive pulmonary disease, and sinusitis. nitric oxide (no) might play an important role in host defense through its potent antiviral properties. previous studies have shown that hrv infection in human subjects increased nasal epithelial expression of type 2 nitric oxide synthase (nos2), an isoform of the enzyme that produces no.	2004	15100676
in vitro activity of expanded-spectrum pyridazinyl oxime ethers related to pirodavir: novel capsid-binding inhibitors with potent antipicornavirus activity.	picornaviruses (pv) include human rhinovirus (hrv), the primary cause of the common cold, and the enteroviruses (ev), which cause serious diseases such as poliomyelitis, meningoencephalitis, and systemic neonatal disease. although no compounds for pv infections have been approved in the united states, pirodavir was one of the most promising capsid-binding compounds to show efficacy in human clinical trials for chemoprophylaxis of the common cold. susceptibility to hydrolysis precluded its use as ...	2004	15105133
gaining target access for deoxyribozymes.	antisense oligonucleotides and ribozymes have been used widely to regulate gene expression by targeting mrnas in a sequence-specific manner. long rnas, however, are highly structured molecules. thus, up to 90% of putative cleavage sites have been shown to be inaccessible to classical rna based ribozymes or dnazymes. here, we report the use of modified nucleotides to overcome barriers raised by internal structures of the target rna. in our attempt to cleave a broad range of picornavirus rnas, we ...	2004	15136038
mode of action of 2-furylmercury chloride, an anti-rhinovirus compound.	2-furylmercury chloride (2-fmc), an organic mercury derivative, has been found to inhibit the replication of all tested human rhinovirus (hrv) serotypes belonging to the antiviral group b and a limited number of hrv serotypes belonging to the antiviral group a. the mechanism of action of 2-fmc was tested against hrv-2 (antiviral group b, minor receptor group), and compared with an antiviral compound for which the viral target was already determined (enviroxime). 2-fmc was found to bind reversibl ...	2004	15168800
identification of the human rhinovirus serotype 1a binding site on the murine low-density lipoprotein receptor by using human-mouse receptor chimeras.	human rhinovirus serotype 1a (hrv1a) binds more strongly to the mouse low-density lipoprotein receptor (ldlr) than to the human homologue (m. reithmayer, a. reischl, l. snyers, and d. blaas, j. virol. 76:6957-6965, 2002). here, we used this fact to determine the binding site of hrv1a by replacing selected ligand binding modules of the human receptor with the corresponding ligand binding modules of the mouse receptor. the chimeric proteins were expressed in mouse fibroblasts deficient in endogeno ...	2004	15194751
twelve receptor molecules attach per viral particle of human rhinovirus serotype 2 via multiple modules.	the crystallographic t = 1 (pseudo t = 3) icosahedral symmetry of the human rhinovirus capsid dictates the presence of 60 identical, symmetry related surface structures that are available for antibody and receptor binding. x-ray crystallography has shown that 60 individual very-low density lipoprotein receptor (vldlr) modules bind to hrv2. their arrangement around the fivefold axes of the virion suggested that tandem oligomers of such modules could attach simultaneously to symmetry-related sites ...	2004	15196928
amplicon sequencing and improved detection of human rhinovirus in respiratory samples.	improved knowledge of the genotypic characteristics of human rhinovirus (hrv) is required, as are nucleic detection assays with the capacity to overcome both the similarities between members of the family picornaviridae and the wide diversity of different hrv serotypes. the goal of the present study was to investigate the variability and the genotypic diversity of clinical strains circulating in the community. since most reverse transcription (rt)-pcr assays available cannot differentiate hrv fr ...	2004	15243084
dynamic force microscopy for imaging of viruses under physiological conditions.	dynamic force microscopy (dfm) allows imaging of the structure and the assessment of the function of biological specimens in their physiological environment. in dfm, the cantilever is oscillated at a given frequency and touches the sample only at the end of its downward movement. accordingly, the problem of lateral forces displacing or even destroying bio-molecules is virtually inexistent as the contact time and friction forces are reduced. here, we describe the use of dfm in studies of human rh ...	2004	15243650
fluorescence labeling of human rhinovirus capsid and analysis by capillary electrophoresis.	the capsid of human rhinovirus serotype 2, consisting of four viral proteins, was fluorescence-labeled with fluorescein isothiocyanate and analyzed by capillary electrophoresis using uv and laser-induced fluorescence detection. heat denaturation, proteolytic digestion, and receptor binding were applied for confirmation of the identity of the peak with the labeled virus. incomplete derivatization with the fluorophore preserved the affinity of the virus for its receptor, indicating that its cell e ...	2004	15253660
sequence analysis of human rhinoviruses in the rna-dependent rna polymerase coding region reveals large within-species variation.	human rhinoviruses (hrvs; family picornaviridae), the most frequent causative agents of respiratory infections, comprise more than 100 distinct serotypes. according to previous phylogenetic analysis of the vp4/vp2-coding sequences, all but one of the hrv prototype strains distribute between the two established species, human rhinovirus a (hrv-a) and human rhinovirus b (hrv-b). here, partial sequences of the rna-dependent rna polymerase (3d polymerase)-coding gene of 48 hrv prototype strains and ...	2004	15269368
all five cold-shock domains of unr (upstream of n-ras) are required for stimulation of human rhinovirus rna translation.	efficient translation of human rhinovirus-2 (hrv-2) rna from its internal ribosome entry site (ires) depends on the presence of cellular trans-acting factors upstream of n-ras (unr) and polypyrimidine-tract-binding protein. unr contains five cold-shock domains (csds) and is predicted to act as an rna chaperone, allowing the hrv-2 ires to attain the correct conformation for ribosome binding. to investigate the role of each of the csds in ires-dependent translation, five unr mutants, each harbouri ...	2004	15269369
poliovirus receptor cd155-targeted oncolysis of glioma.	cell adhesion molecules of the immunoglobulin superfamily are aberrantly expressed in malignant glioma. amongst these, the human poliovirus receptor cd155 provides a molecular target for therapeutic intervention with oncolytic poliovirus recombinants. poliovirus has been genetically modified through insertion of regulatory sequences derived from human rhinovirus type 2 to selectively replicate within and destroy cancerous cells. efficacious oncolysis mediated by poliovirus derivatives depends on ...	2004	15279713
the crystal structure of the rna-dependent rna polymerase from human rhinovirus: a dual function target for common cold antiviral therapy.	human rhinoviruses (hrv), the predominant members of the picornaviridae family of positive-strand rna viruses, are the major causative agents of the common cold. given the lack of effective treatments for rhinoviral infections, virally encoded proteins have become attractive therapeutic targets. the hrv genome encodes an rna-dependent rna polymerase (rdrp) denoted 3dpol, which is responsible for replicating the viral genome and for synthesizing a protein primer used in the replication. here the ...	2004	15296746
enterovirus 68 is associated with respiratory illness and shares biological features with both the enteroviruses and the rhinoviruses.	enterovirus (ev) 68 was originally isolated in california in 1962 from four children with respiratory illness. since that time, reports of ev68 isolation have been very uncommon. between 1989 and 2003, 12 additional ev68 clinical isolates were identified and characterized, all of which were obtained from respiratory specimens of patients with respiratory tract illnesses. no ev68 isolates from enteric specimens have been identified from these same laboratories. these recent isolates, as well as t ...	2004	15302951
high-resolution structure of a picornaviral internal cis-acting rna replication element (cre).	picornaviruses constitute a medically important family of rna viruses in which genome replication critically depends on a small rna element, the cis-acting replication element (cre), that templates 3d(pol) polymerase-catalyzed uridylylation of the protein primer for rna synthesis, vpg. we report the solution structure of the 33-nt cre of human rhinovirus 14 under solution conditions optimal for uridylylation in vitro. the cre adopts a stem-loop conformation with an extended duplex stem supportin ...	2004	15314212
inhibitors of 3c cysteine proteinases from picornaviridae.	the picornaviridae are among the smallest icosahedral positive-sense single stranded rna containing viruses known, and comprise one of the largest and most important families of human and animal pathogens. the hepatitis a virus (hav) and human rhinovirus (hrv) are important pathogens that belong to the picornavirus family. all picornaviruses have a 3c proteinase that processes an initially biosynthesized precursor protein and is crucial for viral maturation and replication. although it is a cyst ...	2004	15320724
discrimination among rhinovirus serotypes for a variant icam-1 receptor molecule.	intercellular adhesion molecule 1 (icam-1) is the cellular receptor for the major group of human rhinovirus serotypes, including human rhinovirus 14 (hrv14) and hrv16. a naturally occurring variant of icam-1, icam-1kilifi, has altered binding characteristics with respect to different hrv serotypes. hrv14 binds to icam-1 only transiently at physiological temperatures but forms a stable complex with icam-1kilifi. conversely, hrv16 forms a stable complex with icam-1 but does not bind to icam-1kilif ...	2004	15331736
rhinovirus 3c protease precursors 3cd and 3cd' localize to the nuclei of infected cells.	human rhinovirus (hrv) 3c protease (3cpro) plays several important roles in the virus replication cycle. this enzyme cleaves the viral polyprotein at discrete sites to produce mature viral proteins and also inhibits cellular rna transcription. it is not clear, however, whether the observed transcriptional shutoff activities are due to 3cpro itself or to 3cpro-containing precursors, and where 3cpro exerts its effects within infected cells. to address these questions hela cells were infected with ...	2004	15448360
structural and virological studies of the stages of virus replication that are affected by antirhinovirus compounds.	pleconaril is a broad-spectrum antirhinovirus and antienterovirus compound that binds into a hydrophobic pocket within viral protein 1, stabilizing the capsid and resulting in the inhibition of cell attachment and rna uncoating. when crystals of human rhinovirus 16 (hrv16) and hrv14 are incubated with pleconaril, drug occupancy in the binding pocket is lower than when pleconaril is introduced during assembly prior to crystallization. this effect is far more marked in hrv16 than in hrv14 and is m ...	2004	15452226
recent advances in rhinovirus therapeutics.	human rhinoviruses are the major causative agents of the common cold. because there are greater than 100 viral serotypes, little immunological protection is afforded to humans by prior rhinovirus exposure, which accounts for the high incidence of infection. in most cases, rhinovirus leads to a short self-limiting illness. however, for asthmatics, the elderly and immunocompromised patients, rhinovirus infection can lead to life-threatening complications. this has spurred a consistent effort over ...	2004	15578974
labeling of capsid proteins and genomic rna of human rhinovirus with two different fluorescent dyes for selective detection by capillary electrophoresis.	during uncoating of human rhinoviruses, the innermost capsid protein vp4 and the genomic rna are released from the viral protein shell. this process gives rise to subviral particles that are composed of the remaining three capsid proteins vp1, vp2, and vp3. the process is believed to take place in a sequential manner in that first vp4 is expelled resulting in a-particles sedimenting at 135s followed by the rna resulting in b-particles sedimenting at 80s. aiming at ultimately analyzing this proce ...	2004	15595880
targacept active conformation search: a new method for predicting the conformation of a ligand bound to its protein target.	targacept active conformation search (tacs) is a novel variation of well-established three-dimensional quantitative structure--activity relationship methodologies that seeks to determine probable conformation(s) of ligands bound to their protein targets. a combination of affinity or activity data and energetically accessible conformational ensembles, each conformer described by three-dimensional (3-d) sensitive descriptors, forms the basis of the tacs data model. recursive pruning is used to red ...	2004	15615532
nitric oxide and the common cold.	the common cold is a clinical syndrome triggered by a variety of viral pathogens, but rhinoviruses are the most frequent cause. complications of such infections include sinusitis, otitis media, and exacerbations of asthma and chronic obstructive lung disease. there is growing interest in host innate defence responses that may regulate the severity of viral responses. we will review recent evidence that nitric oxide is an important contributor to the host response during colds.	2005	15643342
biopolymer stochastic moments. i. modeling human rhinovirus cellular recognition with protein surface electrostatic moments.	stochastic moments may be applied as molecular descriptors in quantitative structure-activity relationship (qsar) studies for small molecules (h. gonzález-dìaz et al., journal of molecular modeling, 2002, vol. 8, pp. 237-245; 2003, vol. 9, pp. 395-407). however, applications in the field of biopolymers are less known. recently, the march-inside approach has been generalized to encode structural features of proteins and other biopolymers (h. gonzález-dáaz et al., bioinformatics, 2003, vol. 19, pp ...	2005	15648087
conservation of amino acids in human rhinovirus 3c protease correlates with broad-spectrum antiviral activity of rupintrivir, a novel human rhinovirus 3c protease inhibitor.	the picornavirus 3c protease is required for the majority of proteolytic cleavages that occur during the viral life cycle. comparisons of published amino acid sequences from 6 human rhinoviruses (hrv) and 20 human enteroviruses (hev) show considerable variability in the 3c protease-coding region but strict conservation of the catalytic triad residues. rupintrivir (formerly ag7088) is an irreversible inhibitor of hrv 3c protease with potent in vitro activity against all hrv serotypes (48 of 48), ...	2005	15673742
phylogenetic analysis of human rhinovirus capsid protein vp1 and 2a protease coding sequences confirms shared genus-like relationships with human enteroviruses.	phylogenetic analysis of the capsid protein vp1 coding sequences of all 101 human rhinovirus (hrv) prototype strains revealed two major genetic clusters, similar to that of the previously reported vp4/vp2 coding sequences, representing the established two species, human rhinovirus a (hrv-a) and human rhinovirus b (hrv-b). pairwise nucleotide identities varied from 61 to 98 % within and from 46 to 55 % between the two hrv species. interserotypic sequence identities in both hrv species were more v ...	2005	15722530
novel [(biphenyloxy)propyl]isoxazole derivatives for inhibition of human rhinovirus 2 and coxsackievirus b3 replication.	during this study, novel biphenyl derivatives were synthesized and tested for antiviral activity.	2005	15743897
human airway epithelial cells produce ip-10 (cxcl10) in vitro and in vivo upon rhinovirus infection.	human rhinovirus (hrv) infections trigger exacerbations of asthma and chronic obstructive pulmonary disease (copd) and are associated with lymphocytic infiltration of the airways. we demonstrate that infection of primary cultures of human airway epithelial cells, or of the beas-2b human bronchial epithelial cell line, with human rhinovirus type 16 (hrv-16) induces expression of cxcl10 [ifn-gamma-inducible protein 10 (ip-10)], a ligand for the cxcr3 receptor found on activated type 1 t lymphocyte ...	2005	15764644
2-ethoxybenzoxazole as a bioisosteric replacement of an ethyl benzoate group in a human rhinovirus (hrv) capsid binder.	a series of pyridazinylpiperidinyl capsid-binding compounds with novel bicyclic substituents were synthesized and screened against human rhinovirus (hrv). several 2-alkoxy- and 2-alkylthio-benzoxazole and benzothiazole derivatives showed excellent anti-hrv activity. when tested against a panel of 16 representative hrv types the 2-ethoxybenzoxazole derivative 13 was found to have superior hrv activity (median ec(50) 3.88ng/ml) to known capsid-binders pleconaril and pirodavir. compound 13 illustra ...	2005	15808466
crystal structure of complete rhinovirus rna polymerase suggests front loading of protein primer.	picornaviruses utilize virally encoded rna polymerase and a uridylylated protein primer to ensure replication of the entire viral genome. the molecular details of this mechanism are not well understood due to the lack of structural information. we report the crystal structure of human rhinovirus 16 3d rna-dependent rna polymerase (hrv16 3dpol) at a 2.4-a resolution, representing the first complete polymerase structure from the picornaviridae family. hrv16 3dpol shares the canonical features of o ...	2005	15596823
opening of size-selective pores in endosomes during human rhinovirus serotype 2 in vivo uncoating monitored by single-organelle flow analysis.	the effect of virus uncoating on endosome integrity during the early steps in viral infection was investigated. using fluid-phase uptake of 10- and 70-kda dextrans labeled with a ph-dependent fluorophore (fluorescein isothiocyanate [fitc]) and a ph-independent fluorophore (cyanine 5 [cy5]), we determined the phs of labeled compartments in intact hela cells by fluorescence-activated cell sorting analysis. subsequently, the number and ph of fluorescent endosomes in cell homogenates were determined ...	2005	15613329
regulation of the cell-cycle-dependent internal ribosome entry site of the pitslre protein kinase: roles of unr (upstream of n-ras) protein and phosphorylated translation initiation factor eif-2alpha.	the pitslre kinases belong to the large family of cyclin-dependent protein kinases. their function has been related to cell-cycle regulation, splicing and apoptosis. we have previously shown that the open reading frame of the p110(pitslre) transcript contains an ires (internal ribosome entry site) that allows the expression of a smaller p58(pitslre) isoform during the g2/m stage of the cell cycle. in the present study we investigated further the role of cis- and trans-acting factors in the regul ...	2005	15330758
an authentic 3' noncoding region is necessary for efficient poliovirus replication.	picornavirus rna replication involves the specific synthesis of negative-strand intermediates followed by an accumulation of positive-strand viral rna in the presence of a multitude of cellular mrnas. previously, in an effort to identify cis-acting elements required for initiation of negative-strand rna synthesis, we deleted the entire 3' noncoding regions from human rhinovirus and poliovirus genomic rnas. these deletion mutation transcripts displayed a severe delay in rna accumulation following ...	2005	16140772
visualization of single receptor molecules bound to human rhinovirus under physiological conditions.	dynamic force microscopy (dfm) was used to image human rhinovirus hrv2 alone and complexed with single receptor molecules under near physiological conditions. specific and site-directed immobilization of hrv2 on a model cell membrane resulted in a crystalline arrangement of virus particles with hexagonal symmetry and 35 nm spacing. high-resolution imaging of the virus capsid revealed about 20 resolvable structural features with 3 nm diameters; this finding is in agreement with protrusions seen b ...	2005	16154082
pharmacophore modeling, docking, and principal component analysis based clustering: combined computer-assisted approaches to identify new inhibitors of the human rhinovirus coat protein.	the development and application of a sophisticated virtual screening and selection protocol to identify potential, novel inhibitors of the human rhinovirus coat protein employing various computer-assisted strategies are described. a large commercially available database of compounds was screened using a highly selective, structure-based pharmacophore model generated with the program catalyst. a docking study and a principal component analysis were carried out within the software package cerius a ...	2005	16190752
[detection of human rhinovirus genes from clinical sample by one-step rt-pcr].	human rhinovirus (hrv) is the most common respiratory pathogen, which causes not only acute respiratory infection and community acquired pneumonitis in children, but also asthma episode and deterioration. however, the detection of respiratory pathogen, which mainly focuses on respiratory syncytial virus, influenzaviruses a and b, parainfluenza viruses 1-3 and adenoviruses, does not include hrv yet by now in china. the absence of detection method limits the clinical understanding of hrv pathogeni ...	2005	16191293
insights into the genetic basis for natural phenotypic resistance of human rhinoviruses to pleconaril.	recent phylogenetic analyses of the deduced amino acid sequence of the major viral capsid protein (vp1) of all human rhinovirus (hrv) serotypes revealed two distinct species within the genus: species a (75 serotypes) and species b (25 serotypes). pleconaril is a novel capsid inhibitor of hrvs. all 75 species a serotypes and 18 of the 25 species b serotypes are susceptible to inhibition by pleconaril in cell culture. the seven resistant serotypes are hrv-4, -5, -42, -84, -93, -97 and -99. we were ...	2005	16199099
inhibition of polyprotein processing and rna replication of human rhinovirus by pyrrolidine dithiocarbamate involves metal ions.	pyrrolidine dithiocarbamate (pdtc) is an antiviral compound that was shown to inhibit the replication of human rhinoviruses (hrvs), poliovirus, and influenza virus. to elucidate the mechanism of pdtc, the effects on the individual steps of the infection cycle of hrv were investigated. pdtc did not interfere with receptor binding or internalization by receptor mediated endocytosis of hrv2 particles into hela cells. but we demonstrate that the processing of the viral polyprotein was prevented by p ...	2005	16254325
[rhinovirus diseases: pathogenesis, diagnostics and treatment].	contemporary data on human rhinovirus diseases and their pathogenesis are presented. special attention is paid to complications which may be caused by rhinovirus infections in allergy-susceptible patients. furthermore, approaches to the diagnostics and treatment of rhinovirus diseases are described. in particular, the advantages of molecular methods for the diagnostics of rhinovirus infection (based on the pcr) in comparison with cultural and immunochemical methods are pointed out. new investiga ...	2005	16279553
a mutation in the first ligand-binding repeat of the human very-low-density lipoprotein receptor results in high-affinity binding of the single v1 module to human rhinovirus 2.	minor group human rhinoviruses (hrvs) bind members of the low-density lipoprotein receptor family for cell entry. the ligand-binding domains of these membrane proteins are composed of various numbers of direct repeats of about 40 amino acids in length. residues involved in binding of module 3 (v3) of the very-low-density lipoprotein receptor (vldlr) to hrv2 have been identified by x-ray crystallography (n. verdaguer, i. fita, m. reithmayer, r. moser, and d. blaas, nat. struct. mol. biol. 11:429- ...	2005	16282473
amino acid changes in proteins 2b and 3a mediate rhinovirus type 39 growth in mouse cells.	many steps of viral replication are dependent on the interaction of viral proteins with host cell components. to identify rhinovirus proteins involved in such interactions, human rhinovirus 39 (hrv39), a virus unable to replicate in mouse cells, was adapted to efficient growth in mouse cells producing the viral receptor icam-1 (icam-l cells). amino acid changes were identified in the 2b and 3a proteins of the adapted virus, rv39/l. changes in 2b were sufficient to permit viral growth in mouse ce ...	2005	15827151
apoptotic events induced by human rhinovirus infection.	hela and 16hbe14o(-) bronchial epithelium cells infected with human rhinovirus serotype 14 (hrv14) were found to exhibit typical apoptotic morphological alterations, such as cell contraction and nuclear condensation. these events coincided with high-molecular-weight dna fragmentation, activation of caspase-9 and caspase-3 and poly(adp-ribose) polymerase cleavage. caspase activation was preceded by cytochrome c translocation from the mitochondria to the cytoplasm, indicating that apoptosis caused ...	2005	15831950
cryptic promoter activity in the dna sequence corresponding to the pim-1 5'-utr.	the serine/threonine kinase pim-1 mrna contains a long and g/c rich 5'-untranslated region (5'-utr). previous work suggested that the pim-1 5'-utr harbors an internal ribosomal entry site (ires) allowing for internal initiation of translation. however, several previously reported eukaryotic ires elements actually contain cryptic promoter activity. to test whether an ires or a cryptic promoter is present in the pim-1 5'-utr, the 5'-utr was re-examined using stringent test procedures. our results ...	2005	15843687
human rhinovirus type 89 variants use heparan sulfate proteoglycan for cell attachment.	we have previously isolated mutants of the major-group human rhinovirus type 89 that grow in cells deficient in intercellular adhesion molecule 1 (icam-1), the receptor used by the wild-type virus for cell entry [a. reischl, m. reithmayer, g. winsauer, r. moser, i. goesler, and d. blaas., j. virol. 75:9312-9319, 2001]. we now demonstrate that one of these variants utilizes heparan sulfate proteoglycan (hspg) as a cellular receptor. adaptation to icam-1-deficient cells not only resulted in the ne ...	2005	15857982
genetic determinants of cell type-specific poliovirus propagation in hek 293 cells.	the ability of poliovirus to propagate in neuronal cells can be reduced by introducing appropriate nucleotide substitutions into the viral genome. specific mutations scattered throughout the poliovirus genome yielded the live attenuated vaccine strains of poliovirus. neuron-specific propagation deficits of the sabin strains are partially encrypted within a confined region of the internal ribosomal entry site (ires), which carries attenuating point mutations in all three serotypes. recently, high ...	2005	15858012
capsid structure and dynamics of a human rhinovirus probed by hydrogen exchange mass spectrometry.	viral capsids are dynamic protein assemblies surrounding viral genomes. despite the high-resolution structures determined by x-ray crystallography and cryo-electron microscopy, their in-solution structure and dynamics can be probed by hydrogen exchange. we report here using hydrogen exchange combined with protein enzymatic fragmentation and mass spectrometry to determine the capsid structure and dynamics of a human rhinovirus, hrv14. capsid proteins (vp1-4) were labeled with deuterium by incubat ...	2005	15883190
dissociation of an antiviral compound from the internal pocket of human rhinovirus 14 capsid.	win antiviral compounds bind human rhinovirus, as well as enterovirus and parechovirus, in an internal cavity located within the viral protein capsid. access to the buried pocket necessitates deviation from the average viral protein structure identified by crystallography. we investigated the dissociation of win 52084 from the pocket in human rhinovirus 14 by using an adiabatic, biased molecular dynamics simulation method. multiple dissociation trajectories are used to characterize the pathway. ...	2005	15899980
inhibition of cellular protein secretion by picornaviral 3a proteins.	during poliovirus infection, anterograde traffic between the endoplasmic reticulum and the golgi is inhibited due to the action of 3a, an 87 amino acid viral protein. the ability of poliovirus protein 3a to inhibit er-to-golgi traffic is not required for virus growth. instead, we have suggested that the inhibition of host protein secretion, shown to reduce the secretion of interferon-beta, il-6, and il-8 and the expression of both newly synthesized mhc class i and tnf receptor in the plasma memb ...	2005	15914217
in vitro antiviral activity and single-dose pharmacokinetics in humans of a novel, orally bioavailable inhibitor of human rhinovirus 3c protease.	(e)-(s)-4-((s)-2-{3-[(5-methyl-isoxazole-3-carbonyl)-amino]-2-oxo-2h-pyridin-1-yl}-pent-4-ynoylamino)-5-((s)-2-oxo-pyrrolidin-3-yl)-pent-2-enoic acid ethyl ester (compound 1) is a novel, irreversible inhibitor of human rhinovirus (hrv) 3c protease {inactivation rate constant (kobs/[i]) of 223,000 m-1s-1}. in cell-based assays, compound 1 was active against all hrv serotypes (35 of 35), hrv clinical isolates (5 of 5), and related picornaviruses (8 of 8) tested with mean 50% effective concentratio ...	2005	15917520
the minor receptor group of human rhinovirus (hrv) includes hrv23 and hrv25, but the presence of a lysine in the vp1 hi loop is not sufficient for receptor binding.	like all 10 minor receptor group human rhinoviruses (hrvs), hrv23 and hrv25, previously classified as major group viruses, are neutralized by maltose binding protein (mbp)-v33333 (a soluble recombinant concatemer of five copies of repeat 3 of the very-low-density lipoprotein receptor fused to mbp), bind to low-density lipoprotein receptor in virus overlay blots, and replicate in intercellular adhesion molecule 1 (icam-1)-negative cos-7 cells. from phylogenetic analysis of capsid protein vp1-codi ...	2005	15919894
human rhinovirus 3c protease: generation of pharmacophore models for peptidic and nonpeptidic inhibitors and their application in virtual screening.	three-dimensional pharmacophore models for peptidic and small organic nonpeptidic inhibitors of the human rhinovirus 3c protease were generated in a structure-based as well as in a ligand-based approach, using the software package catalyst. the inhibitors possess an electrophilic moiety, often a michael acceptor function, which covalently binds to a cysteine in the active site of the enzyme. since this process presents the key step for virus inactivation, the creation of a new function in cataly ...	2005	15921461
the role of p38 mapk in rhinovirus-induced monocyte chemoattractant protein-1 production by monocytic-lineage cells.	viral respiratory infections are a major cause of asthma exacerbations and can contribute to the pathogenesis of asthma. major group human rhinovirus enters cells by binding to the cell surface molecule icam-1 that is present on epithelial and monocytic lineage cells. the focus of the resulting viral infection is in bronchial epithelia. however, previous studies of the cytokine dysregulation that follows rhinovirus infection have implicated monocytic lineage cells in establishing the inflammator ...	2005	15944313
neutralization of a common cold virus by concatemers of the third ligand binding module of the vldl-receptor strongly depends on the number of modules.	concatemers of various numbers of the third ligand binding repeat of human very-low density lipoprotein receptor arranged in tandem were fused to maltose-binding protein and expressed as soluble polypeptides. these artificial receptors protected hela cells against infection with human rhinovirus serotype 2 (hrv2) to a degree that strongly increased with the number of repeats present; maximal protection was seen for the pentameric concatemer (mbp-v33333). this v3 pentamer neutralized hrv2 more ef ...	2005	15950998
antiviral activity of engystol: an in vitro analysis.	to study the effects of the homeopathic preparation engystol (biologische heilmittel heel gmbh, baden-baden, germany) on a panel [corrected] of human pathogenic viruses in vitro.	2005	16296918
ion transport blockers inhibit human rhinovirus 2 release.	picornavirus replication causes leakage of cytoplasmic k+ and an influx of na+ and ca2+. in this study, we have explored the possibility that a blockade of ca2+ and na+ influx would reduce rhinovirus production and/or release. the ca2+-channel blockers, verapamil and diltiazem, as well as the blocker of na+/h+ exchange and the epithelial na+ channel, eipa, inhibited both virus production and release. the effect on virus release was more pronounced than the effect on production, thus raising the ...	2005	16054245
peripheral genotype-phenotype correlations in asian indians with type 2 diabetes mellitus.	a genome-wide scan of gene expression in leucocytes in asian indians with type 2 diabetes was performed and correlated with their known phenotype.	2005	16121806
lack of respiratory and contact sensitizing potential of the intranasal antiviral drug candidate rupintrivir (ag7088): a weight-of-the-evidence evaluation.	rupintrivir, also known as ag7088, is a small molecule 3c protease inhibitor designed to target human rhinovirus as a potential intranasal treatment for the common cold. the ability of rupintrivir to induce both respiratory and contact hypersensitivity responses was evaluated using a weight of the evidence approach. a local lymph node assay (llna) in mice evaluating concentrations of rupintrivir up to 50% in dimethylformamide showed no evidence of sensitizing capability. an irritation study cond ...	2005	18958666
effects of picornavirus 3a proteins on protein transport and gbf1-dependent cop-i recruitment.	the 3a protein of the coxsackievirus b3 (cvb3), an enterovirus that belongs to the family of the picornaviruses, inhibits endoplasmic reticulum-to-golgi transport. recently, we elucidated the underlying mechanism by showing that cvb3 3a interferes with adp-ribosylation factor 1 (arf1)-dependent cop-i recruitment to membranes by binding and inhibiting the function of gbf1, a guanine nucleotide exchange factor that is required for the activation of arf1 (e. wessels et al., dev. cell 11:191-201, 20 ...	2006	17005635
nonneutralizing human rhinovirus serotype 2-specific monoclonal antibody 2g2 attaches to the region that undergoes the most dramatic changes upon release of the viral rna.	the monoclonal antibody 2g2 has been used extensively for detection and quantification of structural changes of human rhinovirus serotype 2 during infection. it recognizes exclusively a and b subviral particles, not native virus. we have elucidated the basis of this selectivity by determining the footprint of 2g2. since viral escape mutants obviously cannot be obtained, the structures of complexes between fab fragments of 2g2 and 80s subviral b particles were determined by cryoelectron microscop ...	2006	17005641
defining residues involved in human rhinovirus 2a proteinase substrate recognition.	the 2a proteinase (2a(pro)) of human rhinoviruses (hrvs) initiates proteolytic processing by cleaving between the c-terminus of vp1 and its own n-terminus. it subsequently cleaves the host protein eif4gi. hrv2 and hrv14 2a(pro) cleave at iitta gpsd and diksy glgp on their respective polyproteins. the hrv2 2a(pro) cleavage site on eif4gi is tlstr *gppr. we show that hrv2 2a(pro) can self-process at the eif4gi cleavage sequence whereas hrv14 2a(pro) cannot, due to the presence of the arginine re ...	2006	17007846
human rhinovirus induces robust ip-10 release by monocytic cells, which is independent of viral replication but linked to type i interferon receptor ligation and stat1 activation.	human rhinovirus (hrv)-induced respiratory infections are associated with elevated levels of ifn-gamma-inducible protein 10 (ip-10), which is an enhancer of t lymphocyte chemotaxis and correlates with symptom severity and t lymphocyte number. increased ip-10 expression is exhibited by airway epithelial cells following ex vivo hrv challenge and requires intracellular viral replication; however, there are conflicting reports regarding the necessity of type i ifn receptor ligation for ip-10 express ...	2006	17020930
alignment of capsid protein vp1 sequences of all human rhinovirus prototype strains: conserved motifs and functional domains.	an alignment was made of the deduced amino acid sequences of the entire capsid protein vp1 of all human rhinovirus (hrv) prototype strains to examine conserved motifs in the primary structure. a set of previously proposed crucially important amino acids in the footprints of the two known receptor molecules was not conserved in a receptor group-specific way. in contrast, vp1 and vp3 amino acids in the minor receptor-group strains corresponding to most of the predicted icam-1 footprint definitely ...	2006	16361425
structure and dynamics of coxsackievirus b4 2a proteinase, an enyzme involved in the etiology of heart disease.	the 2a proteinases (2a(pro)) from the picornavirus family are multifunctional cysteine proteinases that perform essential roles during viral replication, involving viral polyprotein self-processing and shutting down host cell protein synthesis through cleavage of the eukaryotic initiation factor 4g (eif4g) proteins. coxsackievirus b4 (cvb4) 2a(pro) also cleaves heart muscle dystrophin, leading to cytoskeletal dysfunction and the symptoms of human acquired dilated cardiomyopathy. we have determin ...	2006	16415022
age-dependent poliovirus replication in the mouse central nervous system is determined by internal ribosome entry site-mediated translation.	mouse cells are not permissive for the replication of human rhinovirus type 2 (hrv2). to determine the role of the hrv2 internal ribosome entry site (ires) in determining species specificity, a recombinant poliovirus (p1/hrv2) was constructed by substituting the poliovirus ires with the ires from hrv2. this recombinant virus replicated in all human and murine cell lines examined, demonstrating that the hrv2 ires does not limit viral replication in transformed murine cells. p1/hrv2 replicated in ...	2006	16501069
influence of detergent additives on mobility of native and subviral rhinovirus particles in capillary electrophoresis.	the electrophoretic properties of two human rhinovirus (hrv) serotypes, hrv2 and hrv14, their subviral particles, and their capsid proteins were investigated by ce using borate buffer, ph 8.3, as bge and three different detergents as additives. in addition, the influence of modification of the capsid with an amine reactive fluorescent dye, cy3.5, on migration in the electric field was assessed. we found that the reproducibility of the electrophoretic results was decisively dependent on the prese ...	2006	16523456
cell-type-specific repression of internal ribosome entry site activity by double-stranded rna-binding protein 76.	translation of picornavirus plus-strand rna genomes occurs via internal ribosomal entry at highly structured 5' untranslated regions. in addition to canonical translation factors, translation rate is likely influenced by supplementary host and viral trans-acting factors. we previously reported that insertion of a heterologous human rhinovirus type 2 internal ribosomal entry site (ires) into the poliovirus (pv) genome, generating the chimeric virus pv-ripo, selectively abrogates viral translation ...	2006	16537583
rhinoviral infections activate p38map-kinases via membrane rafts and rhoa.	rhinoviral infections belong to the most frequent human infections characterized by common cold, chronic bronchitis, exacerbations of asthma, otitis media and sinusitis. here, we define molecular mechanisms that mediate infections of human epithelial cells with human rhinovirus strain 14 (rv14). we demonstrate that rv14 activates p38-mapkinase (p38-k) in a biphasic time course. early stimulation of p38-k by rv14 was observed a few minutes after initiation of the infection, while the late increas ...	2006	16543732
s-nitrosothiols regulate cell-surface ph buffering by airway epithelial cells during the human immune response to rhinovirus.	human rhinovirus infection is a common trigger for asthma exacerbations. asthma exacerbations and rhinovirus infections are both associated with markedly decreased ph and ammonium levels in exhaled breath condensates. this observation is thought to be related, in part, to decreased activity of airway epithelial glutaminase. we studied whether direct rhinovirus infection and/or the host immune response to the infection decreased airway epithelial cell surface ph in vitro. interferon-gamma and tum ...	2006	16603595
human rhinovirus attenuates the type i interferon response by disrupting activation of interferon regulatory factor 3.	the type i interferon (ifn) response requires the coordinated activation of the latent transcription factors nf-kappab, interferon regulatory factor 3 (irf-3), and atf-2, which in turn activate transcription from the ifn-beta promoter. synthesis and subsequent secretion of ifn-beta activate the jak/stat signaling pathway, resulting in the transcriptional induction of the full spectrum of antiviral gene products. we utilized high-density microarrays to examine the transcriptional response to rhin ...	2006	16641293
evidence for functional protein interactions required for poliovirus rna replication.	poliovirus protein 2c contains a predicted n-terminal amphipathic helix that mediates association of the protein with the membranes of the viral rna replication complex. a chimeric virus that contains sequences encoding the 18-residue core from the orthologous amphipathic helix from human rhinovirus type 14 (hrv14) was constructed. the chimeric virus exhibited defects in viral rna replication and produced minute plaques on hela cell monolayers. large plaque variants that contained mutations with ...	2006	16699013
capillary electrophoresis of affinity complexes between subviral 80s particles of human rhinovirus and monoclonal antibody 2g2.	human rhinoviruses (hrvs), the main etiologic agents of the common cold, transform into subviral b- or 80s particles (they sediment at 80s upon sucrose density gradient centrifugation) during infection and, in vitro, upon exposure to a temperature between 50 and 56 degrees c. with respect to the native virion they lack the genomic rna and the viral capsid protein vp4. 80s particles are unstable and easily disintegrate into their components, vp1, vp2, and vp3 in buffers containing sds. however, t ...	2006	16732623