Publications

the checkpoint saccharomyces cerevisiae rad9 protein contains a tandem tudor domain that recognizes dna.	dna damage checkpoints are signal transduction pathways that are activated after genotoxic insults to protect genomic integrity. at the site of dna damage, 'mediator' proteins are in charge of recruiting 'signal transducers' to molecules 'sensing' the damage. budding yeast rad9, fission yeast crb2 and metazoan 53bp1 are presented as mediators involved in the activation of checkpoint kinases. here we show that, despite low sequence conservation, rad9 exhibits a tandem tudor domain structurally cl ...	2007	17726056
wide-scale analysis of human functional transcription factor binding reveals a strong bias towards the transcription start site.	transcription factors (tf) regulate expression by binding to specific dna sequences. a binding event is functional when it affects gene expression. functionality of a binding site is reflected in conservation of the binding sequence during evolution and in over represented binding in gene groups with coherent biological functions. functionality is governed by several parameters such as the tf-dna binding strength, distance of the binding site from the transcription start site (tss), dna packing, ...	2007	17726537
ubiquitin-dependent proteolytic control of sumo conjugates.	posttranslational protein modification with small ubiquitin-related modifier (sumo) is an important regulatory mechanism implicated in many cellular processes, including several of biomedical relevance. we report that inhibition of the proteasome leads to accumulation of proteins that are simultaneously conjugated to both sumo and ubiquitin in yeast and in human cells. a similar accumulation of such conjugates was detected in saccharomyces cerevisiae ubc4 ubc5 cells as well as in mutants lacking ...	2007	17728242
effect of suramin on the human pathogen candida albicans: implications on the fungal development and virulence.	candida albicans is an opportunistic pathogen that is of growing medical importance because it causes superficial, mucosal and systemic infections in susceptible individuals. here, the effect of suramin, a polysulfonated naphthylurea derivative, on c. albicans development and virulence was evaluated. firstly, it was demonstrated that suramin (500 microm) arrested its growth, showing a fungicidal action dependent on cell number. suramin treatment caused profound changes in the yeast ultrastructur ...	2007	17760875
sumo-targeted ubiquitin ligases in genome stability.	we identify the sumo-targeted ubiquitin ligase (stubl) family of proteins and propose that stubls selectively ubiquitinate sumoylated proteins and proteins that contain sumo-like domains (slds). stubl recruitment to sumoylated/sld proteins is mediated by tandem sumo interaction motifs (sims) within the stubls n-terminus. stubl-mediated ubiquitination maintains sumoylation pathway homeostasis by promoting target protein desumoylation and/or degradation. thus, stubls establish a novel mode of comm ...	2007	17762865
identification and functional characterization of ask/dbf4, a novel cell survival gene in cutaneous melanoma with prognostic relevance.	malignant melanoma is one of the most aggressive and invasive metastatic tumors derived from melanocytes that have undergone malignant transformation by acquisition of genetic and epigenetic alterations. oligonucleotide microarray-based screening of distinct stages in the tumor progression model of cutaneous melanoma identified ask/dbf4, as a novel determinant for melanoma development. quantitative real-time polymerase chain reaction-based confirmation of ask/dbf4 on a series of benign nevi, dys ...	2007	17768177
the morphogenetic regulator czf1p is a dna-binding protein that regulates white opaque switching in candida albicans.	czf1p has been demonstrated to regulate the switch between the yeast-cell morphology and filamentous morphologies of the human fungal pathogen candida albicans. the predicted amino acid sequence of czf1p contains a zinc-cluster motif similar to the dna-binding domains of proteins such as saccharomyces cerevisiae gal4p, suggesting that czf1p is a dna-binding protein. czf1p also demonstrates genetic interaction and a two-hybrid interaction with a second regulator of c. albicans cellular morphology ...	2007	17768232
mrna trafficking in fungi.	fungal growth depends on active transport of macromolecules along the actin and/or microtubule cytoskeleton. thereby, molecular cargo such as proteins, lipids, and mrnas is targeted to defined subcellular regions. active transport and localisation of mrnas mediate localised translation so that protein synthesis occurs where protein function is required. in saccharomyces cerevisiae, actomyosin-dependent mrna trafficking participates in polar growth, asymmetric cell division, targeting of membrane ...	2007	17768642
mutation in mouse hei10, an e3 ubiquitin ligase, disrupts meiotic crossing over.	crossing over during meiotic prophase i is required for sexual reproduction in mice and contributes to genome-wide genetic diversity. here we report on the characterization of an n-ethyl-n-nitrosourea-induced, recessive allele called mei4, which causes sterility in both sexes owing to meiotic defects. in mutant spermatocytes, chromosomes fail to congress properly at the metaphase plate, leading to arrest and apoptosis before the first meiotic division. mutant oocytes have a similar chromosomal p ...	2007	17784788
the u3 small nucleolar ribonucleoprotein component imp4p is a telomeric dna-binding protein.	imp4p is a component of u3 snornp (small nucleolar ribonucleoprotein) involved in the maturation of 18s rrna. we have shown that imp4p interacts with cdc13p, a single-stranded telomere-binding protein involved in telomere maintenance. to understand the role of imp4p in telomeres, we purified recombinant imp4p protein and tested its binding activity towards telomeric dna using electrophoretic mobility-shift assays. our results showed that imp4p bound specifically to single-stranded telomeric dna ...	2007	17803460
enzymatic and hemolytic activities of candida dubliniensis strains.	candida dubliniensis is an opportunistic yeast that has been recovered from several body sites in many populations; it is most often recovered from the oral cavities of human immunodeficiency virus-infected patients. although extensive studies on epidemiology and phylogeny of c. dubliniensis have been performed, little is known about virulence factors such as exoenzymatic and hemolytic activities. in this study we compared proteinase, hyaluronidase, chondroitin sulphatase and hemolytic activitie ...	2007	17823746
generation of a prophylactic melanoma vaccine using whole recombinant yeast expressing mart-1.	malignant melanoma is a potentially deadly form of skin cancer and people at high-risk of developing melanoma will benefit from effective preventive intervention. yeast can be used as an efficient vehicle of antigen loading and immunostimulation. saccharomyces cerevisiae is not pathogenic to humans and can be easily engineered to express specific antigens. in this study, we have developed a melanoma vaccine using a yeast-based platform expressing a full-length melanocyte/melanoma protein to inve ...	2007	17845213
mining a yeast library for brain endothelial cell-binding antibodies.	we describe the use of yeast surface display for the identification of antibodies that bind the plasma membranes of living cells. yeast panning with a nonimmune human single-chain antibody library identified 34 unique lead antibodies that bind (k(d) = 82 +/- 15 nm) and in some cases internalize into rat brain endothelial cells. in addition, we used a new yeast display immunoprecipitation procedure for initial characterization of the cognate antigens.	2007	17206151
a 40.7 kda rpp30/rpp1 homologue is a protein subunit of dictyostelium discoideum rnase p holoenzyme.	rnase p is an essential and ubiquitous endonuclease that mediates the maturation of the 5' ends of all precursor trna molecules. the holoenzyme from dictyostelium discoideum possesses rna and protein subunits essential for activity, but the exact composition of the ribonucleoprotein complex is still under investigation. bioinformatic analysis of d. discoideum genome identified seven open reading frames encoding candidate rnase p protein subunits. the gene named drpp30 encodes a protein with a pr ...	2007	17207566
identification and characterization of putative tumor suppressor ngb, a gtp-binding protein that interacts with the neurofibromatosis 2 protein.	mutations of the neurofibromatosis 2 (nf2) tumor suppressor gene have frequently been detected not only in schwannomas and other central nervous system tumors of nf2 patients but also in their sporadic counterparts and malignant tumors unrelated to the nf2 syndrome such as malignant mesothelioma, indicating a broader role for the nf2 gene in human tumorigenesis. however, the mechanisms by which the nf2 product, merlin or schwannomin, is regulated and controls cell proliferation remain elusive. h ...	2007	17210637
the effect of genetic background on the function of saccharomyces cerevisiae mlh1 alleles that correspond to hnpcc missense mutations.	germline mutations in the dna mismatch repair (mmr) gene mlh1 are associated with a large percentage of hereditary non-polyposis colorectal cancers. there are approximately 250 known human mutations in mlh1. of these, one-third are missense variants that are often difficult to characterize with regards to pathogenicity. we analysed 28 alleles of baker's yeast mlh1 that correspond to non-truncating human mutant alleles listed in online hnpcc databases, 13 of which had not been previously studied ...	2007	17210669
cloning and characterization of the ribosomal protein l3 (rpl3) gene family from triticum aestivum.	plant pathogenic fungi of the genus fusarium can cause severe diseases on small grain cereals and maize. the contamination of harvested grain with fusarium mycotoxins is a threat to human and animal health. in wheat production of the toxin deoxynivalenol (don), which inhibits eukaryotic protein biosynthesis, is a virulence factor of fusarium, and resistance against don is considered to be part of fusarium resistance. previously, single amino acid changes in rpl3 (ribosomal protein l3) conferring ...	2007	17216491
a systems approach to measuring the binding energy landscapes of transcription factors.	a major goal of systems biology is to predict the function of biological networks. although network topologies have been successfully determined in many cases, the quantitative parameters governing these networks generally have not. measuring affinities of molecular interactions in high-throughput format remains problematic, especially for transient and low-affinity interactions. we describe a high-throughput microfluidic platform that measures such properties on the basis of mechanical trapping ...	2007	17218526
a general amphipathic alpha-helical motif for sensing membrane curvature.	the golgi-associated protein arfgap1 has an unusual membrane-adsorbing amphipathic alpha-helix: its polar face is weakly charged, containing mainly serine and threonine residues. we show that this feature explains the specificity of arfgap1 for curved versus flat lipid membranes. we built an algorithm to identify other potential amphipathic alpha-helices rich in serine and threonine residues in protein databases. among the identified sequences, we show that three act as membrane curvature sensor ...	2007	17220896
relationship between mrna stability and length: an old question with a new twist.	the half-life of individual mrna plays a central role in controlling the level of gene expression. however, the determinants of mrna stability have not yet been well defined. most previous studies suggest that mrna length does not affect its stability. here, we show significant negative correlations between mrna length and stability in human and escherichia coli, but not in saccharomyces cerevisiae or bacillus subtilis. this finding suggests the possibility that endonucleolytic attacks by rna en ...	2007	17221301
atx1-like chaperones and their cognate p-type atpases: copper-binding and transfer.	copper is an essential yet toxic metal ion. to satisfy cellular requirements, while, at the same time, minimizing toxicity, complex systems of copper trafficking have evolved in all cell types. the best conserved and most widely distributed of these involve atx1-like chaperones and p(1b)-type atpase transporters. here, we discuss current understanding of how these chaperones bind cu(i) and transfer it to the atx1-like n-terminal domains of their cognate transporter.	2007	17225061
breast cancer-associated gene 3 (bca3) is a novel rac1-interacting protein.	bca3 was identified in a yeast two-hybrid screen as a novel rac1-interacting partner in osteoclasts. bca3 binds directly to rac and, in vivo, binds gtp-rac but not gdp-rac. perinuclear co-localization of bca3 and rac1 is observed in csf-1-treated osteoclasts. overexpression of bca3 attenuates csf-1-induced cell spreading. we conclude that bca3 regulates csf-1-dependent rac activation.	2007	17227220
polyadenylation and decay of 26s rrna as part of nicotiana tabacum response to cadmium.	in contrast to mrnas, ribosomal rnas are generally not considered to be polyadenylated. only a few recent reports describe non-abundant polyadenylated rrna-related transcripts that have been detected and characterized in yeast and in human cells. here we depict the phenomenon of 26s rrna polyadenylation and degradation that was observed in shoots of nicotiana tabaccum plants grown in the presence of cadmium. fragments corresponding to 26s rrna were identified using suppression subtractive hybrid ...	2007	18066401
[micirobial diversity and screening of antitumor activity of actinomycete strains from saline and alkaline environments in the qinghai province, p. r. china].	soil and sediment samples were collected from saline and alkaline soils or lakes in the qinghai province, northwestern china. 145 actinomycete strains were isolated using glucose-peptone-yeast extract agar (gpy) and isp medium 2 agar supplemented with 1.0 - 3.0 mol/l nacl at ph 7.5 - 10. the antitumor activities in vitro of the fermentation broth extracts from the 145 test strains were detected in 6 human tumor cell lines (gastric cancer gxf251l, lung cancer lxfl529l, mammary cancer maxf401nl, m ...	2007	18062244
using homologous recombination to manipulate the genome of human somatic cells.		2007	18059634
engineering the properties of d-amino acid oxidases by a rational and a directed evolution approach.	d-amino acid oxidase (daao) is a fad-containing flavoprotein that dehydrogenates the d-isomer of amino acids to the corresponding imino acids, coupled with the reduction of fad. the cofactor then reoxidizes on molecular oxygen and the imino acid hydrolyzes spontaneously to the alpha-keto acid and ammonia. in vitro daao displays broad substrate specificity, acting on several neutral and basic d-amino acids: the most efficient substrates are amino acids with hydrophobic side chains. d-aspartic aci ...	2007	18220846
brewer's/baker's yeast (saccharomyces cerevisiae) and preventive medicine: part i.	yeast is the term generally applied to a unicellular fungus, and there are hundreds of species now identified. one of the most notable and well-known species of yeast in health and wellness is known as saccharomyces cerevisiae, which is also known by its more common names, brewer's yeast or baker's yeast. it is usually grown on hops or another substrate similar to the plant utilized in the beer-making industry, after which it is harvested and killed. the final product is generally half composed ...	2007	18217542
outer mitochondrial membrane protein degradation by the proteasome.	protein turnover is used for regulatory processes and to eliminate superfluous, denatured or chemically inactivated polypeptides. mitochondrial proteins may be particularly susceptible to damage induced by reactive oxygen species and several pathways of mitochondrial proteolysis have been illuminated. however, in contrast to matrix and inner mitochondrial membrane protein degradation, little is known about the turnover of integral outer mitochondrial membrane (omm) proteins or the mechanisms inv ...	2007	18074628
isolation and characterization of human antibodies targeting human aspartyl (asparaginyl) beta-hydroxylase.	over-expression of the enzyme human aspartyl (asparaginyl) beta-hydroxylase (haah) has been detected in a variety of cancers. it is proposed that upon cellular transformation, haah is overexpressed and translocated to the tumor cell surface, rendering it a specific surface antigen for tumor cells. in this work, twelve human single-chain fv fragments (scfv) against haah were isolated from a human non-immune scfv library displayed on the surface of yeast. five of the twelve were reformatted as hum ...	2007	18334751
pp2a-bgamma subunit and kcnq2 k+ channels in bipolar disorder.	many bipolar affective disorder (bd) susceptibility loci have been identified but the molecular mechanisms responsible for the disease remain to be elucidated. in the locus 4p16, several candidate genes were identified but none of them was definitively shown to be associated with bd. in this region, the ppp2r2c gene encodes the bgamma-regulatory subunit of the protein phosphatase 2a (pp2a-bgamma). first, we identified, in two different populations, single nucleotide polymorphisms and risk haplot ...	2007	16733521
novel function of human adam15 disintegrin-like domain and its derivatives in platelet aggregation.	a disintegrin and metalloproteinase (adam) proteins are a family of multifunctional proteins containing disintegrin and metalloproteinase domains that perform both adhesive and proteolytic functions in cell-cell and cell-matrix interactions. adam15 is unique among these proteins in having an arg-gly-asp (rgd) motif in its disintegrin-like domain. this motif is known to interact with the integrin alphaiibbeta3 on platelets.	2007	16797059
differential effects of rgs proteins on g alpha(q) and g alpha(11) activity.	heterotrimeric g proteins play a pivotal role in gpcr signalling; they link receptors to intracellular effectors and their inactivation by rgs proteins is a key factor in resetting the pathway following stimulation. the precise gpcr:g protein:rgs combination determines the nature and duration of the response. investigating the activity of particular combinations is difficult in cells which contain multiples of each component. we have therefore utilised a previously characterised yeast system to ...	2007	16843638
inactive full-length p53 mutants lacking dominant wild-type p53 inhibition highlight loss of heterozygosity as an important aspect of p53 status in human cancers.	over 1000 different mutants of the tumor suppressor protein p53 with one amino acid change in the core domain have been reported in human cancers. in mouse knock-in models, two frequent mutants displayed loss of wild-type (wt) p53 function, inhibition of wt p53 and wt p53-independent gain of function. the remaining mutants have been systematically characterized for loss of wt p53 function, but not other phenotypes. we report the concomitant assessment of loss of function and interference with wt ...	2007	16861262
occurrence of estrogenic effects in sewage and industrial wastewaters in beijing, china.	estrogenic potencies of the effluents or water samples from wastewater treatment plants (wwtps), industries and hospitals and some receiving rivers in beijing city were estimated by using a human estrogen receptor recombinant yeast assay. estrogenic activity of industrial wastewaters was found to range from 0.1 to 13.3 ng eeq/l and decreased to the range of 0.03-1.6 ng eeq/l after treatment. estrogenic activity in wwtp influent ranged from 0.3 to 1.7 ng eeq/l and decreased to the range of 0.05-0 ...	2007	16872730
human rps19, the gene mutated in diamond-blackfan anemia, encodes a ribosomal protein required for the maturation of 40s ribosomal subunits.	diamond-blackfan anemia (dba) typically presents with red blood cell aplasia that usually manifests in the first year of life. the only gene currently known to be mutated in dba encodes ribosomal protein s19 (rps19). previous studies have shown that the yeast rps19 protein is required for a specific step in the maturation of 40s ribosomal subunits. our objective here was to determine whether the human rps19 protein functions at a similar step in 40s subunit maturation. studies where rps19 expres ...	2007	16990592
estrogenic activity associated with organochlorine compounds in fish extracts from european mountain lakes.	fish muscle extracts from ten european mountain lakes were analyzed for organochlorine compounds (ocs) and estrogenic activity, the latter by a recombinant yeast assay based on the human estrogen receptor. seventy percent of the samples showed estrogenic activity above detection limits and a subset of five samples showed estrogenic activities, equivalent to more than 10,000 pg/g of estradiol. these highly estrogenic samples occurred in two lakes, velké hinçovo in the tatra mountains and redon in ...	2007	16997436
biochemistry of eukaryotic homologous recombination.	the biochemistry of eukaryotic homologous recombination caught fire with the discovery that rad51 is the eukaryotic homolog of the bacterial reca and t4 uvsx proteins; and this field is still hot. the core reaction of homologous recombination, homology search and dna strand invasion, along with the proteins catalyzing it, are conserved throughout evolution in principle. however, the increased complexity of eukaryotic genomes and the diversity of eukaryotic cell biology pose additional challenges ...	2007	21552479
development of a recombinant yeast assay to detect ah-receptor ligands.	endocrine systems of humans and animals are disturbed by dioxin-like compounds, which are ligands of the aryl hydrocarbon receptor (ahr). it is important to determine the accumulation of dioxin-like compounds in the environment for maintenance of human health. in this study, we developed a new method for screening ligands of the ahr using a yeast hybrid system. reporter genes constructed by the insertion of dioxin response elements were integrated into his and lacz yeast genomes. then yeast was ...	2006	20021027
tcc1p, a novel protein containing the tetratricopeptide repeat motif, interacts with tup1p to regulate morphological transition and virulence in candida albicans.	the transcriptional factor catup1p represses many genes involved in intracellular processes, including the yeast-hypha transition, in the human fungal pathogen candida albicans. using tandem affinity purification technology, we identified a novel protein that interacts with catup1p, named tcc1p (tup1p complex component). tcc1p is a c. albicans-specific protein with a 736-amino-acid polypeptide with four tetratricopeptide repeat (tpr) motifs in the n-terminal portion. tcc1p formed a protein compl ...	2006	16998076
compact, universal dna microarrays to comprehensively determine transcription-factor binding site specificities.	transcription factors (tfs) interact with specific dna regulatory sequences to control gene expression throughout myriad cellular processes. however, the dna binding specificities of only a small fraction of tfs are sufficiently characterized to predict the sequences that they can and cannot bind. we present a maximally compact, synthetic dna sequence design for protein binding microarray (pbm) experiments that represents all possible dna sequence variants of a given length k (that is, all 'k-me ...	2006	16998473
a gene-specific requirement for fact during transcription is related to the chromatin organization of the transcribed region.	the fact complex stimulates transcription elongation on nucleosomal templates. in vivo experiments also involve fact in the reassembly of nucleosomes traversed by rna polymerase ii. since several features of chromatin organization vary throughout the genome, we wondered whether fact is equally required for all genes. we show in this study that the in vivo depletion of spt16, one of the subunits of saccharomyces cerevisiae fact, strongly affects transcription of three genes, gal1, pho5, and kluyv ...	2006	17000768
modulation of abeta42 low-n oligomerization using a novel yeast reporter system.	while traditional models of alzheimer's disease focused on large fibrillar deposits of the abeta42 amyloid peptide in the brain, recent work suggests that the major pathogenic effects may be attributed to sds-stable oligomers of abeta42. these abeta42 oligomers represent a rational target for therapeutic intervention, yet factors governing their assembly are poorly understood.	2006	17002801
functional interaction between human herpesvirus 6 immediate-early 2 protein and ubiquitin-conjugating enzyme 9 in the absence of sumoylation.	the immediate-early 2 (ie2) protein of human herpesvirus 6 is a potent transactivator of cellular and viral promoters. to better understand the biology of ie2, we generated a lexa-ie2 fusion protein and screened, using the yeast two-hybrid system, a jurkat t-cell cdna library for proteins that could interact with ie2. the most frequently isolated ie2-interacting protein was the human ubiquitin-conjugating enzyme 9 (ubc9), a protein involved in the small ubiquitin-like modifier (sumo) conjugation ...	2006	17005699
the mrg domain of human mrg15 uses a shallow hydrophobic pocket to interact with the n-terminal region of pam14.	mrg15 is a transcription factor expressed in a variety of human tissues, and its orthologs have been found in many other eukaryotes which constitute the mrg protein family. it plays a vital role in embryonic development and cell proliferation, and is involved in cellular senescence. the c-terminal part of mrg15 forms a conserved mrg domain which is involved in interactions with the tumor suppressor protein retinoblastoma and a nucleoprotein pam14 during transcriptional regulation. we report here ...	2006	17008723
measuring "free" iron levels in caenorhabditis elegans using low-temperature fe(iii) electron paramagnetic resonance spectroscopy.	oxidative stress, caused by free radicals within the body, has been associated with the process of aging and many human diseases. because free radicals, in particular superoxide, are difficult to measure, an alternative indirect method for measuring oxidative stress levels has been used successfully in escherichia coli and yeast. this method is based on a proposed connection between elevated superoxide levels and release of iron from solvent-exposed [4fe-4s] enzyme clusters that eventually leads ...	2006	17010298
nonsense-mediated mrna decay: target genes and functional diversification of effectors.	recent genome-wide identification of nonsense-mediated mrna decay (nmd) targets in yeast, fruitfly and human cells has provided insight into the biological functions and evolution of this mrna quality control mechanism, revealing that nmd post-transcriptionally regulates an important fraction of the transcriptome. nmd targets are associated with a broad range of biological processes, but most of these targets are not encoded by orthologous genes across different species. yeast and fruitfly nmd e ...	2006	17010613
secretory expression of synthetic human fas ligand extracellular domain gene in pichia pastoris: influences of tag addition and n-glycosylation site deletion, and development of a purification method.	human fas ligand is a medically important membrane glycoprotein that induces the apoptosis of harmful cells. a new secretory expression and purification method was devised for the production of a large amount of recombinant human fas ligand extracellular domain (hfaslecd) by pichia pastoris. the expression plasmid containing a synthetic hfaslecd gene designed using yeast optimal codons was constructed for the secretion of hfaslecd. the secreted product exhibited the specific binding activity tow ...	2006	17011210
mtorc2 caught in a sinful akt.	the target of rapamycin (tor), a central controller of cell growth, is found in two distinct, highly conserved multiprotein complexes. three recent papers in cell (jacinto et al., 2006), developmental cell (shiota et al., 2006; this issue), and current biology (frias et al., 2006) shed light on mtor complex 2 (mtorc2) composition and in vivo function. an important new finding is that mtorc2 determines akt/pkb substrate specificity rather than absolute activity.	2006	17011481
long-range downstream enhancers are essential for pax6 expression.	pax6 is a developmental control gene with an essential role in development of the eye, brain and pancreas. pax6, as many other developmental regulators, depends on a substantial number of cis-regulatory elements in addition to its promoters for correct spatiotemporal and quantitative expression. here we report on our analysis of a set of mice transgenic for a modified yeast artificial chromosome carrying the human pax6 locus. in this 420 kb yac a taugfp-ires-neomycin reporter cassette has been i ...	2006	17014839
an artificially constructed de novo human chromosome behaves almost identically to its natural counterpart during metaphase and anaphase in living cells.	human artificial chromosomes (hacs) are promising reagents for the analysis of chromosome function. while hacs are maintained stably, the segregation mechanisms of hacs have not been investigated in detail. to analyze hacs in living cells, we integrated 256 copies of the lac operator into a precursor yeast artificial chromosome (yac) containing alpha-satellite dna and generated green fluorescent protein (gfp)-tagged hacs in ht1080 cells expressing a gfp-lac repressor fusion protein. time-lapse a ...	2006	17015481
the common phospholipid-binding activity of the n-terminal domains of pex1 and vcp/p97.	pex1 is a type ii aaa-atpase that is indispensable for biogenesis and maintenance of the peroxisome, an organelle responsible for the primary metabolism of lipids, such as beta-oxidation and lipid biosynthesis. recently, we demonstrated a striking structural similarity between its n-terminal domain and those of other membrane-related aaa-atpases, such as valosine-containing protein (p97). the n-terminal domain of valosine-containing protein serves as an interface to its adaptor proteins p47 and ...	2006	17018057
mechanism of actin filament turnover by severing and nucleation at different concentrations of adf/cofilin.	adf/cofilins are key regulators of actin dynamics during cellular motility, yet their precise role and mechanism of action are shrouded in ambiguity. direct observation of actin filaments by evanescent wave microscopy showed that cofilins from fission yeast and human do not increase the rate that pointed ends of actin filaments shorten beyond the rate for adp-actin subunits, but both cofilins inhibit elongation and subunit dissociation at barbed ends. direct observation also showed that cofilins ...	2006	17018289
human papillomavirus type 5 e6 oncoprotein represses the transforming growth factor beta signaling pathway by binding to smad3.	mechanisms of cellular transformation associated with human papillomavirus type 5 (hpv5), which is responsible for skin carcinomas in epidermodysplasia verruciformis (ev) patients, are poorly understood. using a yeast two-hybrid screening and molecular and cellular biology experiments, we found that hpv5 oncoprotein e6 interacts with smad3, a key component in the transforming growth factor beta1 (tgf-beta1) signaling pathway. hpv5 e6 inhibits smad3 transactivation by destabilizing the smad3/smad ...	2006	17020941
estrogenic activity of uv filter mixtures.	uv-absorbing chemicals (uv filters) are widely used for protection against uv radiation in sunscreens and in a variety of cosmetic products and materials. depending on the breadth and factor of uv protection, they are added as single compounds or as a combination thereof. some uv filters have estrogenic activity, but their activity and interactions in mixtures are largely unknown. in this work, we analyzed 8 commonly used uv filters, which are pure or partial heralpha agonists, for their estroge ...	2006	17027055
functional characterization of human and cynomolgus monkey udp-glucuronosyltransferase 1a6 enzymes.	udp-glucuronosyltransferase 1a6 (ugt1a6) is a major isoform in the human liver that glucuronidates numerous drugs, environmental chemicals and endogenous substrates. in this study, human and cynomolgus monkey ugt1a6 cdnas (humugt1a6 and monugt1a6, respectively) were cloned, and the corresponding proteins were heterologously expressed in yeast cells to identify the functions of primate ugt1a6s. the enzymatic properties of ugt1a6 proteins were characterized by the kinetic analysis of serotonin (5- ...	2006	17027947
centromere-proximal crossovers are associated with precocious separation of sister chromatids during meiosis in saccharomyces cerevisiae.	in most organisms, meiotic chromosome segregation is dependent on crossovers (cos), which enable pairs of homologous chromosomes to segregate to opposite poles at meiosis i. in mammals, the majority of meiotic chromosome segregation errors result from a lack of cos between homologs. observations in homo sapiens and drosophila melanogaster have revealed a second class of exceptional events in which a co occurred near the centromere of the missegregated chromosome. we show that in wild-type strain ...	2006	17028345
influence of silicon on cobalt, zinc, and magnesium in baker's yeast, saccharomyces cerevisiae.	silicon (si, as silicate) is involved in numerous important structure and function roles in a wide range of organisms, including man. silicate availability influences metal concentrations within various cell and tissue types, but, as yet, clear mechanisms for such an influence have been discovered only within the diatoms and sponges. in this study, the influence of silicate on the intracellular accumulation of metals was investigated in baker's yeast (saccharomyces cerevisiae). it was found that ...	2006	17028383
identification and characterization of a novel drosophila 3'-phosphoadenosine 5'-phosphosulfate transporter.	sulfation of macromolecules requires the translocation of a high energy form of nucleotide sulfate, i.e. 3'-phosphoadenosine 5'-phosphosulfate (paps), from the cytosol into the golgi apparatus. in this study, we identified a novel drosophila paps transporter gene dpapst2 by conducting data base searches and screening the paps transport activity among the putative nucleotide sugar transporter genes in drosophila. the amino acid sequence of dpapst2 showed 50.5 and 21.5% homology to the human papst ...	2006	16873373
bacello: a balanced subcellular localization predictor.	the knowledge of the subcellular localization of a protein is fundamental for elucidating its function. it is difficult to determine the subcellular location for eukaryotic cells with experimental high-throughput procedures. computational procedures are then needed for annotating the subcellular location of proteins in large scale genomic projects.	2006	16873501
free-energy distribution of binary protein-protein binding suggests cross-species interactome differences.	major advances in large-scale yeast two-hybrid screening have provided a global view of binary protein-protein interactions across species as dissimilar as human, yeast, and bacteria. remarkably, these analyses have revealed that all species studied have a degree distribution of protein-protein binding that is approximately scale-free (varies as a power law) even though their evolutionary divergence times differ by billions of years. the universal power law shows only the surface of the rich inf ...	2006	16861299
recent evolution of the human pathogen cryptococcus neoformans by intervarietal transfer of a 14-gene fragment.	the availability of the whole-genome sequence from the 2 known varieties of the human pathogenic fungus cryptococcus neoformans provides an opportunity to study the relative contribution of divergence and introgression during the process of speciation in a genetically tractable organism. at the genomic level, these varieties are nearly completely syntenic, share approximately 85-90% nucleotide identity, and are believed to have diverged approximately 18 mya. via a comparative genomic approach, w ...	2006	16870684
sugar-dependent1 encodes a patatin domain triacylglycerol lipase that initiates storage oil breakdown in germinating arabidopsis seeds.	triacylglycerol hydrolysis (lipolysis) plays a pivotal role in the life cycle of many plants by providing the carbon skeletons and energy that drive postgerminative growth. despite the physiological importance of this process, the molecular mechanism is unknown. here, a genetic screen has been used to identify arabidopsis thaliana mutants that exhibit a postgerminative growth arrest phenotype, which can be rescued by providing sugar. seventeen sugar-dependent (sdp) mutants were isolated, and six ...	2006	16473965
mdm38 interacts with ribosomes and is a component of the mitochondrial protein export machinery.	saccharomyces cerevisiae mdm38 and ylh47 are homologues of human letm1, a protein implicated in wolf-hirschhorn syndrome. we analyzed the function of mdm38 and ylh47 in yeast mitochondria to gain insight into the role of letm1. we find that mdm38delta mitochondria have reduced amounts of certain mitochondrially encoded proteins and low levels of complex iii and iv and accumulate unassembled atp6 of complex v of the respiratory chain. mdm38 is especially required for efficient transport of atp6 a ...	2006	16476776
ancient origin, functional conservation and fast evolution of dna-dependent rna polymerase iii.	rna polymerase iii contains seventeen subunits in yeasts (saccharomyces cerevisiae and schizosaccharomyces pombe) and in human cells. twelve of them are akin to the core rna polymerase i or ii. the five other are rna polymerase iii-specific and form the functionally distinct groups rpc31-rpc34-rpc82 and rpc37-rpc53. currently sequenced eukaryotic genomes revealed significant homology to these seventeen subunits in fungi, animals, plants and amoebozoans. except for subunit rpc31, this also extend ...	2006	16877568
the effects of glutaredoxin and copper activation pathways on the disulfide and stability of cu,zn superoxide dismutase.	mutations in cu,zn superoxide dismutase (sod1) can cause amyotrophic lateral sclerosis (als) through mechanisms proposed to involve sod1 misfolding, but the intracellular factors that modulate folding and stability of sod1 are largely unknown. by using yeast and mammalian expression systems, we demonstrate here that sod1 stability is governed by post-translational modification factors that target the sod1 disulfide. oxidation of the human sod1 disulfide in vivo was found to involve both the copp ...	2006	16880213
gawky is a component of cytoplasmic mrna processing bodies required for early drosophila development.	in mammalian cells, the gw182 protein localizes to cytoplasmic bodies implicated in the regulation of messenger rna (mrna) stability, translation, and the rna interference pathway. many of these functions have also been assigned to analogous yeast cytoplasmic mrna processing bodies. we have characterized the single drosophila melanogaster homologue of the human gw182 protein family, which we have named gawky (gw). drosophila gw localizes to punctate, cytoplasmic foci in an rna-dependent manner. ...	2006	16880270
jab1 mediates cytoplasmic localization and degradation of west nile virus capsid protein.	the clinical manifestations of west nile virus (wnv), a member of the flavivirus family, include febrile illness, sporadic encephalitis, and paralysis. the capsid (cp) of wnv is thought to participate in these processes by inducing apoptosis through mitochondrial dysfunction and activation of caspase-9 and caspase-3. to further identify the molecular mechanism of the wnv capsid protein (wnvcp), yeast two-hybrid assays were employed using wnv-cp as bait. jab1, the fifth subunit of the cop9 signal ...	2006	16882664
drosophila rtf1 functions in histone methylation, gene expression, and notch signaling.	the rtf1 subunit of the paf1 complex is required for proper monoubiquitination of histone h2b and methylation of histone h3 on lysines 4 (h3k4) and 79 in yeast saccharomyces cerevisiae. using rnai, we examined the role of rtf1 in histone methylation and gene expression in drosophila melanogaster. we show that drosophila rtf1 (drtf1) is required for proper gene expression and development. furthermore, we show that rnai-mediated reduction of drtf1 results in a reduction in histone h3k4 trimethylat ...	2006	16882721
intrinsic disorder is a common feature of hub proteins from four eukaryotic interactomes.	recent proteome-wide screening approaches have provided a wealth of information about interacting proteins in various organisms. to test for a potential association between protein connectivity and the amount of predicted structural disorder, the disorder propensities of proteins with various numbers of interacting partners from four eukaryotic organisms (caenorhabditis elegans, saccharomyces cerevisiae, drosophila melanogaster, and homo sapiens) were investigated. the results of pondr vl-xt dis ...	2006	16884331
copper-dependent interaction of glutaredoxin with the n termini of the copper-atpases (atp7a and atp7b) defective in menkes and wilson diseases.	the p-type atpases affected in menkes and wilson diseases, atp7a and atp7b, respectively, are key copper transporters that regulate copper homeostasis. the n termini of these proteins are critical in regulating their function and activity, and contain six copper-binding motifs mxcxxc. in this study, we describe the identification of glutaredoxin (grx1) as an interacting partner of both atp7a and atp7b, confirmed by yeast two-hybrid technology and by co-immunoprecipitation from mammalian cells. t ...	2006	16884690
identification of gnr1p, a negative regulator of g alpha signalling in schizosaccharomyces pombe, and its complementation by human g beta subunits.	g protein-coupled receptors (gpcrs) are involved in the response of eukaryotic cells to a wide variety of stimuli, traditionally mediating their effects through heterotrimeric g proteins comprised of g alpha, g beta and g gamma subunits. the fission yeast schizosaccharomyces pombe is an established tool for gpcr research, possessing two g alpha-dependent signalling cascades. a complete g alpha beta gamma complex has been characterised for the glucose-sensing pathway, but only the g alpha subunit ...	2006	16884933
fhl2/slim3 decreases cardiomyocyte survival by inhibitory interaction with sphingosine kinase-1.	sphingosine kinase-1 (sk1) is a key enzyme catalyzing the phosphorylation of sphingosine to sphingosine-1-phosphate (s1p). recent studies suggest that sk1, and its product s1p, regulate diverse biological functions, including cell growth, differentiation, proliferation, and apoptosis. s1p may also play an important role in cardiac development and ischemic preconditioning, but the mechanism underlying these effects is not known. using a yeast 2-hybrid screen with sk1 as bait and a cardiac cdna li ...	2006	16888242
using annotated peptide mass spectrum libraries for protein identification.	a system for creating a library of tandem mass spectra annotated with corresponding peptide sequences was described. this system was based on the annotated spectra currently available in the global proteome machine database (gpmdb). the library spectra were created by averaging together spectra that were annotated with the same peptide sequence, sequence modifications, and parent ion charge. the library was constructed so that experimental peptide tandem mass spectra could be compared with those ...	2006	16889405
genetic structure and evolution of the vps25 family, a yeast escrt-ii component.	vps25p is the product of yeast gene vps25 and is found in an endosomal sorting complex required for transport (escrt)-ii, along with vps22p and vps36p. this complex is essential for sorting of ubiquitinated biosynthetic and endosomal cargoes into endosomes.	2006	16889659
interactions between e2f1 and sirt1 regulate apoptotic response to dna damage.	the nicotinamide adenine dinucleotide (nad)-dependent deacetylase sir2 (silent information regulator 2) regulates gene silencing in yeast and promotes lifespan extension during caloric restriction. the mammalian homologue of sir2 (sirt1) regulates p53, nf-kappab and forkhead transcription factors, and is implicated in stress response. this report shows that the cell-cycle and apoptosis regulator e2f1 induces sirt1 expression at the transcriptional level. furthermore, sirt1 binds to e2f1 and inhi ...	2006	16892051
dengue virus ns4b interacts with ns3 and dissociates it from single-stranded rna.	dengue virus, a member of the family flaviviridae of positive-strand rna viruses, has seven non-structural proteins: ns1, ns2a, ns2b, ns3, ns4a, ns4b and ns5. except for enzymic activities contained within ns3 and ns5, the roles of the other proteins in virus replication and pathogenesis are not well defined. in this study, a physical interaction between ns4b and the helicase domain of ns3 was identified by using a yeast two-hybrid assay. this interaction was further confirmed by biochemical pul ...	2006	16894199
heme: a versatile signaling molecule controlling the activities of diverse regulators ranging from transcription factors to map kinases.	heme (iron protoporphyrin ix) is an essential molecule for numerous living organisms. not only does it serve as a prosthetic group in enzymes, it also acts as a signaling molecule that controls diverse molecular and cellular processes ranging from signal transduction to protein complex assembly. deficient heme synthesis or function impacts the hematopoietic, hepatic and nervous systems in humans. recent studies have revealed a series of heme-regulated transcription factors and signal transducers ...	2006	16894358
identification of gamma-aminobutyric acid receptor-interacting factor 1 (trak2) as a trafficking factor for the k+ channel kir2.1.	to identify proteins that regulate potassium channel activity and expression, we performed functional screening of mammalian cdna libraries in yeast that express the mammalian k(+) channel kir2.1. growth of kir2.1-expressing yeast in media with low k(+) concentration is a function of k(+) uptake via kir2.1 channels. therefore, the host strain was transformed with a human cdna library, and cdna clones that rescued growth at low k(+) concentration were selected. one of these clones was identical t ...	2006	16895905
inhibitors of human indoleamine 2,3-dioxygenase identified with a target-based screen in yeast.	indoleamine 2,3-dioxygenase (ido) is a tryptophan degradation enzyme that is emerging as an important drug target. ido is expressed by many human tumors to help them escape immune detection, and it has been implicated in depression and in the formation of senile nuclear cataracts. there is a need for potent and selective ido inhibitors for use in research and as lead compounds for drug development. we show that expression of human ido in a saccharomyces cerevisiae tryptophan auxotroph restricts ...	2006	16897708
molecular dissection of the munc18c/syntaxin4 interaction: implications for regulation of membrane trafficking.	sec1p/munc18 (sm) proteins are believed to play an integral role in vesicle transport through their interaction with snares. different sm proteins have been shown to interact with snares via different mechanisms, leading to the conclusion that their function has diverged. to further explore this notion, in this study, we have examined the molecular interactions between munc18c and its cognate snares as these molecules are ubiquitously expressed in mammals and likely regulate a universal plasma m ...	2006	16899085
elevated expression of human alpha-1 antitrypsin mediated by yeast intron in pichia pastoris.	intron-mediated enhancement has been documented in many cases to involve large positive effect on gene expression. to address this, human alpha-1 antitrypsin (haat) gene was integrated into pichia pastoris with and without a yeast intron generated from the final plasmid pblu-exii-int-exiii and ligated into the ecori/bamhi multiple cloning site of the yeast shuttle vector phil-s1. the chimeric exon-intron complex in the middle of the naturally occurring haat exons ii and iii caused a 23-fold enha ...	2006	16900336
genome comparison using gene ontology (go) with statistical testing.	automated comparison of complete sets of genes encoded in two genomes can provide insight on the genetic basis of differences in biological traits between species. gene ontology (go) is used as a common vocabulary to annotate genes for comparison. current approaches calculate the fold of unweighted or weighted differences between two species at the high-level go functional categories. however, to ensure the reliability of the differences detected, it is important to evaluate their statistical si ...	2006	16901353
molecular cloning and characterization of a novel v-atpase associated protein, dva9.2, from human dendritic cells.	the vacuolar proton-atpase (v-atpase) is a ubiquitous atp-driven h(+) transporter that functions in numerous cell processes. accumulating evidence shows important roles of v-atpase in tumor metastasis and antigen presentation of dendritic cells (dc). a novel v-atpase associated protein, designated as dva9.2 (dendritic cell-derived v-atpase associated protein of 9.2 kda), has been identified from a human dc cdna library by large-scale random sequencing. full length cdna of dva9.2 encodes an 81-re ...	2006	16904702
evaluation of oriental medicinal herbs for estrogenic and antiproliferative activities.	herb extracts commercially used in asia were screened for their estrogenic activity with a recombinant yeast system with both a human estrogen receptor (er) expression plasmid and a reporter plasmid. pueraria lobata (flower) had the highest estrogenic relative potency (rp, 17-estradiol = 1.00) (7.8e-3) (rp for + control), followed by amomum xanthioides (1.3e-3), glycyrrhiza uralensis, zingiber officinale, rheum palmatum, curcuma aromatica, eriobotrya japonica, sophora flavescens, anemarrhena asp ...	2006	16906642
conservative repair of a chromosomal double-strand break by single-strand dna through two steps of annealing.	the repair of chromosomal double-strand breaks (dsbs) is essential to normal cell growth, and homologous recombination is a universal process for dsb repair. we explored dsb repair mechanisms in the yeast saccharomyces cerevisiae using single-strand oligonucleotides with homology to both sides of a dsb. oligonucleotide-directed repair occurred exclusively via rad52- and rad59-mediated single-strand annealing (ssa). even the ssa domain of human rad52 provided partial complementation for a null ra ...	2006	16908537
[molecular dynamics modeling of the substitution of serine for the conservative glycine in the g loop in the yeast cdc28-srm mutant using the crystalline lattice of human kinase cdk2].	two-nanosecond molecular dynamics modeling of the crystalline lattice of an active complex of kinase pt160-cdk2/cyclin a/atp-mg2+ substrate has been performed. the results of modeling indicated that the structures of the nonmutant cdk2 complex and mutant cdk2 complex, which involves the g 16s-cd k2 substitution corresponding to that of yeast, markedly differ, the differences in structural conformations being particularly well pronounced in those regions that play a key role in the functioning of ...	2006	16909847
mammalian clasp1 and clasp2 cooperate to ensure mitotic fidelity by regulating spindle and kinetochore function.	clasps are widely conserved microtubule plus-end-tracking proteins with essential roles in the local regulation of microtubule dynamics. in yeast, drosophila, and xenopus, a single clasp orthologue is present, which is required for mitotic spindle assembly by regulating microtubule dynamics at the kinetochore. in mammals, however, only clasp1 has been directly implicated in cell division, despite the existence of a second paralogue, clasp2, whose mitotic roles remain unknown. here, we show that ...	2006	16914514
molecular characterization of a g protein alpha-subunit-encoding gene from mucor circinelloides.	genes encoding the galpha subunit were cloned from mucor circinelloides, a zygomycete dimorphic fungus. there are at least four genes that encode for galpha subunits, gpa1, gpa2, gpa3, and gpa4. the genes gpa1 and gpa3 were isolated and characterized, and their predicted products showed 36%-67% identity with galpha subunits from diverse fungi. northern blot analysis of gpa3 showed that it is present in spores and constitutively expressed during mycelium development and during yeast-mycelium and ...	2006	16917518
a recombinant transductor-effector system: in vitro study of g inhibitory protein (g-alpha-i1) direct activators.	mutations and altered functionality of the inhibitory subfamily of g proteins (gi) are involved in pathological states. compounds able to activate gi in a receptor-independent manner would be useful to treat these pathological conditions. aimed to study gi direct activation we have reconstituted a recombinant transductor-effector complex cloning both the mammalian galpha(i1) subunit and adenylate cyclase (ac). the myristoylation of galpha, fundamental for interaction with ac, was obtained in the ...	2006	16919234
[structural biology of proteasome].		2006	16922377
[regulators of mammalian cellular autophagy].		2006	16922424
[autophagy and autophagic cell death].		2006	16922431
[structural biology of atg conjugation systems].		2006	16922434
candida albicans biofilms produce antifungal-tolerant persister cells.	fungal pathogens form biofilms that are highly recalcitrant to antimicrobial therapy. the expression of multidrug resistance pumps in young biofilms has been linked to increased resistance to azoles, but this mechanism does not seem to underlie the resistance of mature biofilms that is a model of in vivo infection. the mechanism of drug resistance of mature biofilms remains largely unknown. we report that biofilms formed by the major human pathogen candida albicans exhibited a strikingly biphasi ...	2006	16923951
an atp-binding cassette transporter related to yeast vacuolar scycf1 is important for cd sequestration in chlamydomonas reinhardtii.	we generated a cd-sensitive insertional mutant, cds18, in chlamydomonas reinhardtii and elucidated the deletion of a 10 kb fragment containing the promoter and a portion of the coding region for crmrp2 gene that silenced the transcription of crmrp2 in mutant cds18. the association between crmrp2 and cd sensitivity was confirmed by complementing mutant cds18 with a cloned genomic dna fragment containing the promoter and complete coding sequence for crmrp2. the genomic region and the full-length c ...	2006	16930316
expression and localization of recombinant human edg-1 receptors in pichia pastoris.	the receptor for human endothelial differentiation gene-1 protein (edg-1) was c-terminally tagged with green fluorescent protein and expressed in the methylotrophic yeast, pichia pastoris. edg-1 expression was driven by the highly inducible alcohol oxidase 1 promoter. expression of edg-1 recombinant protein was detected by western blot analysis and confocal microscopy. the recombinant edg-1 receptor protein was located in the plasma membrane. radioligand binding assays demonstrated that the edg- ...	2006	16937248
insights on augmenter of liver regeneration cloning and function.	hepatic stimulator substance (hss) has been referred to as a liver-specific but species non-specific growth factor. gradient purification and sequence analysis of hss protein indicated that it contained the augmenter of liver regeneration (alr), also known as hepatopoietin (hpo). alr, acting as a hepatotrophic growth factor, specifically stimulated proliferation of cultured hepatocytes as well as hepatoma cells in vitro, promoted liver regeneration and recovery of damaged hepatocytes and rescued ...	2006	16937489
isolation of intrinsically active mutants of map kinases via genetic screens in yeast.	intrinsically active variants of a protein are powerful tools for deciphering the specific functions of that protein. since the catalytic activity of such variants is spontaneously active in vivo, they can disclose very accurately biochemical and biological functions of the parental protein. it is particularly important to obtain intrinsically active variants of individual map kinases. this is because in response to extracellular signals, more than one mapk is typically concomitantly activated m ...	2006	16938468
progressive loss of sirt1 with cell cycle withdrawal.	sir2 is an nad+-dependent deacetylase that regulates lifespan in yeast, worms and flies. the mammalian orthologs of sir2 include sirt1 in humans and mice. in this study, we analyzed the level of sirt1 in human lung fibroblasts (imr90) and mouse embryonic fibroblasts (mefs) from mice with normal, accelerated, and delayed aging. sirt1 protein, but not mrna, decreased significantly with serial cell passage in both human and murine cells. mouse sirt1 decreased rapidly in prematurely senescent (p44 t ...	2006	16939484
genetic and chemical rescue of the saccharomyces cerevisiae phenotype induced by mitochondrial dna polymerase mutations associated with progressive external ophthalmoplegia in humans.	the human polg gene encodes the catalytic subunit of mitochondrial dna polymerase gamma (pol gamma). mutations in pol gamma are associated with a spectrum of disease phenotypes including autosomal dominant and recessive forms of progressive external ophthalmoplegia, spino-cerebellar ataxia and epilepsy, and alpers-huttenlocher hepatocerebral poliodystrophy. multiple deletions, or depletion of mtdna in affected tissues, are the molecular hallmarks of pol gamma mutations. to shed light on the path ...	2006	16940310