Publications

dolichol phosphate mannose synthase is required in vivo for glycosyl phosphatidylinositol membrane anchoring, o mannosylation, and n glycosylation of protein in saccharomyces cerevisiae.	glycosyl phosphatidylinositol (gpi) anchoring, n glycosylation, and o mannosylation of protein occur in the rough endoplasmic reticulum and involve transfer of precursor structures that contain mannose. direct genetic evidence is presented that dolichol phosphate mannose (dol-p-man) synthase, which transfers mannose from gdpman to the polyisoprenoid dolichol phosphate, is required in vivo for all three biosynthetic pathways leading to these covalent modifications of protein in yeast cells. tempe ...	1990	2146492
the spermicidal compound nonoxynol-9 increases adhesion of candida species to human epithelial cells in vitro.	all 25 strains of candida spp. tested were able to grow in medium supplemented with 25% nonoxynol-9 in vitro. adhesion of candida spp. to hela cells was found to increase in the presence of 5% nonoxynol-9 (2.2- to 6.6-fold; p less than 0.001) and, to a lesser extent, in 12.5% nonoxynol-9. adhesion of candida strains cultured in medium supplemented with nonoxynol-9 varied, with five of six strains of candida albicans and the single strain of candida tropicalis demonstrating increases of 1.4 to 4. ...	1990	2160437
tissue and species distribution of liver type and tumor type nuclear poly(a) polymerases.	previous studies in this laboratory have identified two distinct nuclear poly(a) polymerases, a 48 kda tumor type enzyme and a 36-38 kda liver type enzyme. to investigate the tissue and species specificity of these enzymes, nuclear extracts were prepared from various rat tissues, pig brain and two human cell lines. these as well as whole cell extract from yeast were probed for the two enzymes by immunoblot analysis using polyclonal anti-tumor poly(a) polymerase antibodies or autoimmune sera whic ...	1990	2162661
phenotypic selection and characterization of mutant alleles of a eukaryotic dna topoisomerase i.	we have developed a simple, effective genetic screen for mutant alleles of eukaryotic dna topoisomerase i that manifest severely depressed or complete loss of enzymatic function. the screen is based on the extreme toxicity of vaccinia topoisomerase expression in the escherichia coli lysogen strain bl21(de3) and is notable for its ease in distinguishing nonsense mutations (that result in truncated proteins) from missense mutations. the power of the method is evinced by our observation that 100% o ...	1990	2163340
the structure of the human gene encoding protein gene product 9.5 (pgp9.5), a neuron-specific ubiquitin c-terminal hydrolase.	database search using a bovine thymus ubiquitin c-terminal hydrolase sequence indicated 54% sequence identity with the abundant human neuron-specific protein gene product 9.5 (pgp9.5), which was then shown to possess the same activity [wilkinson, lee, deshpande, duerksen-hughes, boss & pohl (1989) science 246, 670-673]. a yeast counterpart of the enzyme is also known. the human pgp9.5 gene, described here, spans 10 kb, contains nine exons and displays 5' features some common to many genes and so ...	1990	2163617
killing of coccidioides immitis by hypochlorous acid or monochloramine.	to identify possible explanations for the resistance of coccidioides immitis to killing by human neutrophils, its susceptibility to typical oxidants generated during the neutrophil respiratory burst was compared to the sensitivity of other microbes. when microbial suspensions were exposed to hypochlorous acid, arthroconidia or spherules of c. immitis were killed more slowly than yeast cells of candida (torulopsis) glabrata or staphylococcus aureus. in contrast, exposure to the more lipophilic ox ...	1990	2166156
effect of human interleukin-2 from different sources on lymphocyte and airway beta-adrenoceptor function.	the effect of recombinant human interleukin-2 (rh il-2, genzyme, yeast derived) on the beta-adrenoceptor function of human peripheral blood mononuclear cells (pbmc), guinea pig splenic lymphocytes and isolated guinea pig tracheal spirals was investigated. rh il-2 (genzyme, yeast derived) induces a dose dependent inhibition of the isoprenaline-stimulated camp production in pbmc and splenic lymphocytes after a two hour preincubation period. the inhibition is significant at 0.01 u/ml il-2 and reach ...	1990	2167879
cdna sequence of two distinct pituitary proteins homologous to kex2 and furin gene products: tissue-specific mrnas encoding candidates for pro-hormone processing proteinases.	based on the concept of sequence conservation around the active sites of serine proteinases, polymerase chain reaction applied to mrna amplification allowed us to obtain a 260-bp probe which was used to screen a mouse pituitary cdna library. the primers used derived from the cdna sequence of active sites ser* and asn* of human furin. two cdna sequences were obtained from a number of positive clones. these code for two similar but distinct structures (mpc1 and mpc2), each being homologous to yeas ...	1990	2169760
proton-linked sugar transport systems in bacteria.	the cell membranes of various bacteria contain proton-linked transport systems for d-xylose, l-arabinose, d-galactose, d-glucose, l-rhamnose, l-fucose, lactose, and melibiose. the melibiose transporter of e. coli is linked to both na+ and h+ translocation. the substrate and inhibitor specificities of the monosaccharide transporters are described. by locating, cloning, and sequencing the genes encoding the sugar/h+ transporters in e. coli, the primary sequences of the transport proteins have been ...	1990	2172229
wilms tumor locus on 11p13 defined by multiple cpg island-associated transcripts.	wilms tumor is an embryonal kidney tumor involving complex pathology and genetics. the wilms tumor locus on chromosome 11p13 is defined by the region of overlap of constitutional and tumor-associated deletions. chromosome walking and yeast artificial chromosome (yac) cloning were used to clone and map 850 kilobases of dna. nine cpg islands, constituting a "cpg island archipelago," were identified, including three islands that were not apparent by conventional pulsed-field mapping, and thus were ...	1990	2173146
human alpha-n-acetylgalactosaminidase-molecular cloning, nucleotide sequence, and expression of a full-length cdna. homology with human alpha-galactosidase a suggests evolution from a common ancestral gene.	human alpha-n-acetylgalactosaminidase (alpha-galnac, e.c. 3.2.1.49), the lysosomal glycohydrolase that cleaves alpha-n-acetylgalactosaminyl moieties from glycoconjugates, is encoded by a gene localized to chromosome 22q13----qter. the deficient activity of alpha-galnac is the enzymatic defect in schindler disease, an inherited neuroaxonal dystrophy. to isolate a full-length cdna, the enzyme from human lung was purified to homogeneity, 129 non-overlapping amino acids were determined by microseque ...	1990	2174888
expression of human vitamin d receptor in saccharomyces cerevisiae. purification, properties, and generation of polyclonal antibodies.	we have cloned a cdna encoding the human vitamin d receptor (vdr) into a high copy yeast plasmid controlled transcriptionally by the copper-inducible metallothionein (cup-1) promoter to produce yepv1. introduction of this plasmid in the protease deficient saccharomyces cerevisiae strain bj 3505 and subsequent growth in the presence of copper and 1,25-dihydroxyvitamin d3 leads to the synthesis of intact vdr comprising over 0.5% of total soluble protein. the vdr was purified to near homogeneity fr ...	1990	2174892
heterologous expression of human 5-aminolevulinate dehydratase in saccharomyces cerevisiae.	a cdna coding for human 5-amino-levulinate dehydratase was placed in a yeast expression vector under the control of the gal10 promoter. the resulting multicopy plasmid was then used to transform a yeast mutant which contains a defective hem2 gene coding for 5-aminolevulinate dehydratase. expression of the human cdna was shown in four ways: (1) restoration of normal growth on glycerol/galactose as primary carbon source, (2) decrease in intracellular 5-aminolevulinic acid concentration, (3) restor ...	1990	2178784
structure-function analysis of epidermal growth factor: site directed mutagenesis and nuclear magnetic resonance.	the role of leucine-47 in determining the structure and activity of human epidermal growth factor was examined using site-directed mutagenesis. wild type protein and four variants in which leu47 was replaced by valine, glutamate, aspartate and alanine were produced from yeast. 1h nmr experiments demonstrated that substitution of leu47 had little effect on the protein structure. the observed reduction in receptor binding affinity caused by the substitutions could thus be attributed to perturbatio ...	1990	2178977
the candidate proto-oncogene bcl-3 is related to genes implicated in cell lineage determination and cell cycle control.	a gene, bcl-3, is found on chromosome 19 adjacent to the breakpoints in the translocation t(14;19)(q32;q13.1), which occurs in some cases of chronic lymphocytic leukemia. sequence analysis of the human bcl-3 gene predicts a protein containing seven tandem copies of the swi6/cdc10 motif. this motif was previously identified in yeast genes that regulate events at the start of the cell cycle and in invertebrate transmembrane proteins involved in cell differentiation pathways. expression of bcl-3 in ...	1990	2180580
growth inhibition of cryptococcus neoformans by cultured human monocytes: role of the capsule, opsonins, the culture surface, and cytokines.	despite a presumed critical role of macrophages in the host response to cryptococcal infections, previous studies have failed to show growth inhibition of encapsulated cryptococcus neoformans by human peripheral blood cultured monocyte-derived macrophages (mo-m phi). here, we examined whether mo-m phi could be induced to inhibit growth of an encapsulated strain and an isogenic acapsular mutant strain of c. neoformans. mo-m phi were cultured in microwells, and inhibition was measured by comparing ...	1990	2182538
purification and characterization of human plasminogen activator inhibitor type i expressed in saccharomyces cerevisiae.	the rapidly acting inhibitor of plasminogen activators, pai-1, was produced intracellularly in saccharomyces cerevisiae by using the adh2 promoter to drive the expression of the human pai-1 cdna. approximately 8 mg of human pai-1 was produced per liter of confluent yeast culture. a purification scheme which resulted in 20% recovery of isolated pai-1 from the broken yeast cell homogenate was devised. yeast-derived human pai-1 differs from endothelial-type pai-1 isolated from ht1080 fibrosarcoma c ...	1990	2183723
ercc2: cdna cloning and molecular characterization of a human nucleotide excision repair gene with high homology to yeast rad3.	human ercc2 genomic clones give efficient, stable correction of the nucleotide excision repair defect in uv5 chinese hamster ovary cells. one clone having a breakpoint just 5' of classical promoter elements corrects only transiently, implicating further flanking sequences in stable gene expression. the nucleotide sequences of a cdna clone and genomic flanking regions were determined. the ercc2 translated amino acid sequence has 52% identity (73% homology) with the yeast nucleotide excision repai ...	1990	2184031
human u2 snrna can function in pre-mrna splicing in yeast.	the removal of introns from messenger rna precursors requires five small nuclear rnas (snrnas), contained within ribonucleoprotein particles (snrnps), which complex with the pre-mrna and other associated factors to form the spliceosome. in both yeast and mammals, the u2 snrna base pairs with sequences surrounding the site of lariat formation. binding of u2 snrnp to the highly degenerate branchpoint sequence in mammalian introns is absolutely dependent on an auxiliary protein, u2af, which recogni ...	1990	2185425
subunits shared by eukaryotic nuclear rna polymerases.	rna polymerases i, ii, and iii share three subunits that are immunologically and biochemically indistinguishable. the saccharomyces cerevisiae genes that encode these subunits (rpb5, rpb6, and rpb8) were isolated and sequenced, and their transcriptional start sites were deduced. rpb5 encodes a 25-kd protein, rpb6, an 18-kd protein, and rpb8, a 16-kd protein. these genes are single copy, reside on different chromosomes, and are essential for viability. the fact that the genes are single copy, cor ...	1990	2186966
an autonomously replicating sequence from hela dna shows a similar organization to the yeast ars1 element.	a hela dna fragment, which may function as an anchorage point to the nuclear matrix for human chromosomes 1 and 2, also functions as an autonomously replicating sequence (ars) in the yeast saccharomyces cerevisiae. in the present report we show that this dna fragment contains both bent dna and an a-t rich region which appear to be associated with the ars function. more interestingly, dna sequence analysis shows that the spatial distribution of these features is strikingly similar to that found i ...	1990	2187151
prenyl proteins in eukaryotic cells: a new type of membrane anchor.	recent studies have indicated that eukaryotic cells contain proteins that are post-translationally modified by long-chain, thioether-linked prenyl groups. these proteins include yeast mating factors, ras proteins and nuclear lamins. the modification occurs on a cysteine residue near the c terminus and appears to initiate a set of additional protein modification reactions that promote attachment of the proteins to specific membranes.	1990	2187294
free flow electrophoresis for the purification of proteins: i. zone electrophoresis and isotachophoresis.	the principles and some applications of free flow zone electrophoresis and isotachophoresis are described. the influence of (i) carrier electrolyte conductivity on the migration velocity and (ii) band shape on zone electrophoresis was investigated. the technique was found convenient for studying the effect of ph on the mobility of proteins to create a mobility curve. the purification of alcohol dehydrogenase from a crude yeast extract revealed the separation power of zone electrophoresis for com ...	1990	2187696
enhancement of the gdp-gtp exchange of ras proteins by the carboxyl-terminal domain of scd25.	in saccharomyces cerevisiae, the product of the cdc25 gene controls the ras-mediated production of adenosine 3',5'-monophosphate (camp). in vivo the carboxyl-terminal third of the cdc25 gene product is sufficient for the activation of adenylate cyclase. the 3'-terminal part of scd25, a gene of s. cerevisiae structurally related to cdc25, can suppress the requirement for cdc25. partially purified preparations of the carboxy-terminal domain of the scd25 gene product enhanced the exchange rate of g ...	1990	2188363
the regulation of dna repair during development.	dna repair is important in such phenomena as carcinogenesis and aging. while much is known about dna repair in single-cell systems such as bacteria, yeast, and cultured mammalian cells, it is necessary to examine dna repair in a developmental context in order to completely understand its processes in complex metazoa such as man. we present data to support the notion that proliferating cells from organ systems, tumors, and embryos have a greater dna repair capacity than terminally differentiated, ...	1990	2188652
secretion of n-glycosylated human recombinant interleukin-1 alpha in saccharomyces cerevisiae.	we have expressed fragments of the cdna coding for mature human interleukin-1 alpha (hil-1 alpha) in saccharomyces cerevisiae. mature hil-1 alpha contains one potential n-linked glycosylation site that is not recognized in mammalian cells. translational fusions to either one of three yeast signal sequences resulted in secretion of bioactive, n-glycosylated hil-1 alpha. the extent of glycosylation was significantly reduced using the alpha-factor signal sequence, which itself contains three n-link ...	1990	2189790
invasive infection with saccharomyces cerevisiae: report of three cases and review.	saccharomyces cerevisiae (brewer's or baker's yeast) is a common colonizer of human mucosal surfaces, but its role as a clinically important pathogen has been unclear. we report three cases of life-threatening invasive infection with s. cerevisiae resulting in pneumonia, liver abscess and sepsis, and disseminated infection with cardiac tamponade, respectively. a review of the english-language literature reveals 14 other cases of saccharomyces infection in humans. severe immunosuppression, prolon ...	1990	2193348
cryptococcus neoformans, candida albicans, and other fungi bind specifically to the glycosphingolipid lactosylceramide (gal beta 1-4glc beta 1-1cer), a possible adhesion receptor for yeasts.	the role of glycosphingolipids as adhesion receptors for yeasts was examined. cryptococcus neoformans, candida albicans, and saccharomyces cerevisiae, as well as histoplasma capsulatum and sporotrichum schenckii (in their yeast phases), bound specifically to lactosylceramide (gal beta 1-4glc beta 1-1cer), as measured by overlaying glycosphingolipid chromatograms with 125i-labeled organisms. an unsubstituted galactosyl residue was required for binding, because the yeasts did not bind to glucosylc ...	1990	2194958
cloning and characterization of kluyveromyces lactis sec14, a gene whose product stimulates golgi secretory function in saccharomyces cerevisiae.	the saccharomyces cerevisiae sec14 gene encodes a cytosolic factor that is required for secretory protein movement from the golgi complex. that some conservation of sec14p function may exist was initially suggested by experiments that revealed immunoreactive polypeptides in cell extracts of the divergent yeasts kluyveromyces lactis and schizosaccharomyces pombe. we have cloned and characterized the k. lactis sec14 gene (sec14kl). immunoprecipitation experiments indicated that sec14kl encoded the ...	1990	2198263
amino acid sequences of aspartate aminotransferases: the cytosolic isoenzymes from yeast and from human liver.		1990	2199266
substrate overlap and functional competition between human nucleotide excision repair and escherichia coli photolyase and (a)bc excision nuclease.	human cell free extract prepared by the method of manley et al. (1980) carries out repair synthesis on uv-irradiated dna. removal of pyrimidine dimers by photoreactivation with dna photolyase reduces repair synthesis by about 50%. with excess enzyme in the reaction mixture photolyase reduced the repair signal by the same amount even in the absence of photoreactivating light, presumably by binding to pyrimidine dimers and interfering with the binding of human damage recognition protein. similarly ...	1990	2200513
ligand-specific transactivation of gene expression by a derivative of the human glucocorticoid receptor expressed in yeast.	in this study we have reconstituted transactivation of gene expression by the human glucocorticoid receptor in the yeast, saccharomyces cerevisiae. we have expressed the c-terminal half of the human glucocorticoid receptor (residues 415-777), the smallest derivative that can be expected to function as a ligand-dependent activator of transcription, in yeast cells. the function of the expressed protein has been assayed using a reporter gene consisting of the beta-galactosidase gene from escherichi ...	1990	2203760
a yeast homolog of the human uef stimulates transcription from the adenovirus 2 major late promoter in yeast and in mammalian cell-free systems.	we report the identification and purification of a yeast factor functionally homologous to the human upstream element factor (uefh). although the yeast protein (uefy) has a higher molecular weight than the hela uef (60 kd versus 45 kd) both have identical dna-binding properties: the purified uefy recognizes the adenovirus 2 (ad2) major late promoter upstream element (mlp-ue; from nucleotide -49 to -67) as well as the iva2 upstream element (iva2-ue; from nucleotide -98 to -122) with a higher affi ...	1990	2204028
enzymatic coupling of cholesterol intermediates to a mating pheromone precursor and to the ras protein.	the post-translational processing of the yeast a-mating pheromone precursor, ras proteins, nuclear lamins, and some subunits of trimeric g proteins requires a set of complex modifications at their carboxyl termini. this processing includes three steps: prenylation of a cysteine residue, proteolytic processing, and carboxymethylation. in the yeast saccharomyces cerevisiae, the product of the dpr1-ram1 gene participates in this type of processing. through the use of an in vitro assay with peptide ...	1990	2204115
purification of the upstream element factor of the adenovirus-2 major late promoter from hela and yeast by sequence-specific dna affinity chromatography.	the purification to homogeneity of the adenovirus-2 major late promoter (mlp) upstream element factor (uef), a sequence specific transcription factor, which binds to upstream elements of various class b (ii) genes, is reported. the protein was purified from hela cells and also from the yeast saccharomyces cerevisiae, by using sequence-specific dna affinity chromatography. the human (uefh, 45,000 dalton) and the yeast (uefy, 60,000 dalton) proteins protect the same sequences over the mlp-iva2 int ...	1990	2205616
regulated expression and phosphorylation of the 23-26-kda ras protein in the sponge geodia cydonium.	we have cloned, sequenced and examined the sponge geodia cydonium cdna encoding a protein homologous to ras proteins. the sponge ras protein has a more conserved n-terminal region and a less conserved c-terminal region, especially in comparison to dictyostelium discoideum; the similarity to human c-ha-ras-1 and to saccharomyces cerevisiae is less pronounced. the sponge ras cdna comprises five tag triplets; at the translational level these uag termination codons are suppressed by a gln-trna. the ...	1990	2209606
chromosomal region of the cystic fibrosis gene in yeast artificial chromosomes: a model for human genome mapping.	a general strategy for cloning and mapping large regions of human dna with yeast artificial chromosomes (yac's) is described. it relies on the use of the polymerase chain reaction to detect dna landmarks called sequence-tagged sites (sts's) within yac clones. the method was applied to the region of human chromosome 7 containing the cystic fibrosis (cf) gene. thirty yac clones from this region were analyzed, and a contig map that spans more than 1,500,000 base pairs was assembled. individual yac' ...	1990	2218515
glutaminyl-trna synthetase as a component of the high-molecular weight complex of human aminoacyl-trna synthetases. an immunological study.	the human glutaminyl-trna synthetase is three times larger than the corresponding bacterial and twice as large as the yeast enzyme. it is possible that the additional sequences of the human glutaminyl-trna synthetase are required for the formation of the multienzyme complex which is known to include several of aminoacyl-trna synthetases in mammalian cells. to address this point we prepared antibodies against three regions of the human glutaminyl-trna synthetase, namely against its enzymatically ...	1990	2223884
immunoreactivity of neoglycolipids constructed from oligomannosidic residues of the candida albicans cell wall.	to establish a model to study the immunoreactivity of oligosaccharidic structures from the candida albicans cell wall, we attempted to construct neoglycolipids with these residues by using oligomannosides released after mild acid hydrolysis of the phosphopeptidomannans isolated from yeast forms. from a mixture of manno-oligosaccharides ranging from mannobiose to mannononaose, the structure of a quantitatively major component (mannotriose) was determined to be man (beta 1-2) man (beta 1-2) man al ...	1990	2228224
an amino-terminal c-myc domain required for neoplastic transformation activates transcription.	the product of the c-myc proto-oncogene is a nuclear phosphoprotein whose normal cellular function has not yet been defined. c-myc has a number of biochemical properties, however, that suggest that it may function as a potential regulator of gene transcription. specifically, it is a nuclear dna-binding protein with a short half-life, a high proline content, segments that are rich in glutamine and acidic residues, and a carboxyl-terminal oligomerization domain containing the leucine zipper and he ...	1990	2233723
prenylation of mammalian ras protein in xenopus oocytes.	ras protein requires an intermediate of the cholesterol biosynthetic pathway for posttranslational modification and membrane anchorage. this step is necessary for biological activity. maturation of xenopus laevis oocytes induced by an oncogenic human ras protein can be inhibited by lovastatin or compactin, inhibitors of the synthesis of mevalonate, an intermediate of cholesterol biosynthesis. this inhibition can be overcome by mevalonic acid or farnesyl diphosphate, a cholesterol biosynthetic in ...	1990	2233726
synthesis of a gene for human serum albumin and its expression in saccharomyces cerevisiae.	a 1761 base pairs long artificial gene coding for human serum albumin (hsa) has been prepared by a newly developed synthetic approach, resulting in the largest synthetic gene so far described. oligonucleotides corresponding to only one strand of the hsa gene were prepared by chemical synthesis, while the complementary strand was obtained by a combination of enzymatic and cloning steps. 24 synthetic, 69-85 nucleotides long oligonucleotides covering the major part of the hsa gene (41-1761 nucleoti ...	1990	2235491
streamlined approach to creating yeast artificial chromosome libraries from specialized cell sources.	the study of tumor-specific chromosomal abnormalities has been severely impeded by an inability to link cytogenetic to molecular data. restriction fragment length polymorphism mapping of any particular chromosomal rearrangement to the resolution limit of genetic methodology generates sets of probes that frequently are still too widely spaced to render the rearrangement breakpoints accessible to molecular isolation. the stable propagation of genomic fragments of up to one million base pairs in si ...	1990	2236075
antimitochondrial autoantibodies of primary biliary cirrhosis as a novel probe in the study of 2-oxo acid dehydrogenases in patients with mitochondrial myopathies.	autoantibodies present in the autoimmune disease primary biliary cirrhosis react by immunoblotting with four human skeletal muscle mitochondrial antigens of 70 kda, 52 kda, 50 kda and 45 kda, identified as the lipoate acetyl transferases (e2) of the pyruvate dehydrogenase, component x of e2 pyruvate dehydrogenase, e2 of 2-oxo glutarate dehydrogenase and e2 of branched-chain 2-oxo acid dehydrogenase complexes respectively. these autoantibodies have been employed as a novel probe to study whether ...	1990	2243228
factors involved in specific transcription by mammalian rna polymerase ii: role of transcription factors iia, iid, and iib during formation of a transcription-competent complex.	human transcription factor tfiid, the tata-binding protein, was partially purified to a form capable of associating stably with the tata motif of the adenovirus major late promoter. binding of the human and yeast tfiid to the tata motif was stimulated by tfiia. tfiia is an integral part of a complex capable of binding other transcription factors. a complex formed with human tfiid and tfiia (da complex) was specifically recognized by tfiib. we found that tfiib activity was contained in a single p ...	1990	2247058
second-generation approach to the construction of yeast artificial-chromosome libraries.	we describe an improved method for construction of yeast artificial-chromosome (yac) libraries that contain large inserts of foreign dna. the procedure consists of seven steps: (i) preparation of human dna in agarose beads; (ii) partial digestion of the dna with ecori; (iii) electrophoretic elimination of the smaller partial-digest fragments; (iv) ligation of the ecori fragments with vector arms in molten agarose; (v) hydrolysis of agarose with agarase; (vi) fractionation of the recombinant mole ...	1990	2249850
[the nature of n-terminal signal sequence determines the type of intracellular distribution and effectiveness of export of human growth hormone in saccharomyces cerevisiae].	various n-terminal signal peptides (sp) were tested to investigate a human growth hormone (hgh) synthesis, processing and intracellular sorting in yeast. maximal level of hgh was observed in the case when the mature hgh gene was placed under the control of pho5 promoter. in this case about 90% of hgh was localized in the cytosol, but some portion was trustworthly detected in microsomes and periplasma in spite of the absence of sp. addition of own or pho5 sp resulted in lowering of the synthesis ...	1990	2250679
expression of a human proprotein processing enzyme: correct cleavage of the von willebrand factor precursor at a paired basic amino acid site.	intracellular proteolytic processing of precursor polypeptides is an essential step in the maturation of many proteins, including plasma proteins, hormones, neuropeptides, and growth factors. most frequently, propeptide cleavage occurs after paired basic amino acid residues. to date, no mammalian propeptide processing enzyme with such specificity has been purified or cloned and functionally characterized. we report the isolation and functional expression of a cdna encoding a propeptide-cleaving ...	1990	2251280
physical and immunological characterization of human transcription factor iiia.	human transcription factor iiia (htfiiia), specifically required for transcription of the gene for 5s ribosomal rna has been characterized with respect to some of its physical, immunological and functional properties. tfiiia from hela cells, which selectively binds 5s rna, is a monomer of approximately 35 kda with a stokes' radius of approximately 2.65 nm and a sedimentation coefficient of approximately 2.8 s. these values indicate that the human protein is of rather globular shape and hence div ...	1990	2253613
rapid identification of yeast artificial chromosome clones by matrix pooling and crude lysate pcr.		1990	2263507
cell-cycle aspects of growth and maturation of mammalian oocytes.	in this review, recent data concerning growth and maturation of nonmammalian and mammalian female germ cells are compiled with regard to the increased understanding of somatic cells mitotic cycles, from yeast to human tissues. these data allow us to conclude that growing oocytes of nonvertebrates, lower vertebrates, and mammals resemble somatic cells in the g1 phase of the mitotic cycle in their metabolic and cell cycle behavior. transcriptional and translational activity of growing oocytes and ...	1990	2264998
spermidine biosynthesis in saccharomyces cerevisiae. biosynthesis and processing of a proenzyme form of s-adenosylmethionine decarboxylase.	we have cloned and sequenced the saccharomyces cerevisiae gene for s-adenosylmethionine decarboxylase. this enzyme contains covalently bound pyruvate which is essential for enzymatic activity. we have shown that this enzyme is synthesized as a mr 46,000 proenzyme which is then cleaved post-translationally to form two polypeptide chains: a beta subunit (mr 10,000) from the amino-terminal portion and an alpha subunit (mr 36,000) from the carboxyl-terminal portion. the protein was overexpressed in ...	1990	2266128
a candida albicans homolog of a human cyclophilin gene encodes a peptidyl-prolyl cis-trans isomerase.	a candida albicans cdna and its genomic counterpart were isolated from lambda phage libraries using a human t-cell cyclophilin (cyp) cdna as a hybridization probe. the clones contain a 486-bp open reading frame predicting a 162-amino acid, approx. 18 kda protein which is similar in size to, and which shares 68 and 81% homology with, human t-cell cyp and cytosolic saccharomyces cerevisiae cyp, respectively. northern blots show the presence of a single mrna species of about 800 bp. however, genomi ...	1990	2269432
protein retention in yeast rough endoplasmic reticulum: expression and assembly of human ribophorin i.	the rer retains a specific subset of er proteins, many of which have been shown to participate in the translocation of nascent secretory and membrane proteins. the mechanism of retention of rer specific membrane proteins is unknown. to study this phenomenon in yeast, where no rer-specific membrane proteins have yet been identified, we expressed the human rer-specific protein, ribophorin i. in all mammalian cell types examined, ribophorin i has been shown to be restricted to the membrane of the r ...	1990	2269658
conformational requirement of signal sequences functioning in yeast: circular dichroism and 1h nuclear magnetic resonance studies of synthetic peptides.	recently, we have designed a series of simplified artificial signal sequences and have shown that a proline residue in the signal sequence plays an important role in the secretion of human lysozyme in yeast, presumably by altering the conformation of the signal sequence [yamamoto, y., taniyama, y., & kikuchi, m. (1989) biochemistry 28, 2728-2732]. to elucidate the conformational requirement of the signal sequence in more detail, functional and nonfunctional signal sequences connected to the n-te ...	1990	2271573
a yeast mutant, prp20, altered in mrna metabolism and maintenance of the nuclear structure, is defective in a gene homologous to the human gene rcc1 which is involved in the control of chromosome condensation.	we report on the characterization of the yeast prp20-1 mutant. in this temperature-sensitive mutant, multiple steps of mrna metabolism are affected. the prp20-1 mutant strain showed alterations in mrna steady-state levels, defective mrna splicing and changes in transcription initiation or termination when shifted from the permissive to the non-permissive temperature. in addition, a change in the structure of the nucleus in these cells became apparent. electron microscopy revealed an altered stru ...	1990	2277633
isolation and molecular analysis of the orotidine-5'-phosphate decarboxylase gene (pyrg) of phycomyces blakesleeanus.	the pyrg gene of phycomyces was isolated from a phycomyces genomic library, constructed in the cosmid phs255, by hybridization with a 170 bp fragment of the pyrg gene of aspergillus niger. this fragment includes a consensus sequence found in almost all species in which the orotidine-5'-phosphate decarboxylase (ompdecase) gene has been sequenced. the complete nucleotide sequence of the cloned pyrg gene from phycomyces was determined and the transcription start sites mapped. in the predicted amino ...	1990	2277645
preparation and characterization of monoclonal antibodies to human hexokinase type i.	1. three different immunization protocols and several screening procedures were used to prepare seven mouse monoclonal antibodies to human placenta hexokinase type i. none of these monoclonals were able to recognize the native enzyme but all detected hexokinase when adsorbed onto polystyrene plates or on immunoblots after sds/polyacrylamide-gel electrophoresis. 2. all seven monoclonals recognize the two different subtypes of human hexokinase i equally well. limited tryptic digestion of hexokinas ...	1990	2280763
human xq24-xq28: approaches to mapping with yeast artificial chromosomes.	one hundred twenty-seven yeast strains with artificial chromosomes containing xq24-xqter human dna were obtained starting from a human/hamster somatic cell hybrid. the clones were characterized with respect to their insert size, stability, and representation of a set of xq24-xqter dna probes. the inserts of the clones add up to 19.3 megabase (mb) content, or about 0.4 genomic equivalents of that portion of the x chromosome, with a range of 40-650 kb in individual yacs. eleven clones contained mo ...	1990	2294758
isolation and sequencing of nop1. a yeast gene encoding a nucleolar protein homologous to a human autoimmune antigen.	we have identified the gene for the yeast nucleolar protein p38 and deduced the primary structure of p38 from its sequence. we propose the name nop1 (nucleolar protein 1) for this gene. nop1 encodes a 327 amino acid protein of 34,470 daltons and is flanked by potential promoter and polyadenylation sequences. blot analyses indicate that the mrna transcribed from nop1 is approximately 1.3 kilobases in size and that there is one nop1 gene per haploid genome. the amino-terminal sequence of p38 is ho ...	1990	2298745
reactivation of neutron killed mammalian cells by gamma irradiation: the observations, possible mechanism and implication.	we have observed that combinations of neutron plus gamma ray exposure can significantly increase the colony forming ability of monkey and human cell cultures over the neutron dose alone. the "reactivation" of neutron killed mammalian cells by gamma rays is analogous to observations made in lower eukaryotic organisms and fits the pattern termed "t repair" previously postulated for yeast and protozoans.	1990	2300886
deficient lysosomal carboxypeptidase activity in galactosialidosis.	in the lysosome, the glycosidases neuraminidase (ec 3.2.1.18) and beta-galactosidase (ec 3.2.1.23) are associated to a 52 kda "protective protein" to form a large multi-enzymatic complex. deficient synthesis or inactivation of this protective protein causes galactosialidosis, a lysosomal storage disorder in man in which both neuraminidase and beta-galactosidase activities are deficient. since the protective protein possesses extensive sequence homology with carboxypeptidase y (carb y) and the ke ...	1990	2328002
nuclear proteins from hela cells are immunoreactive with anti-yeast yp20 ras related antibodies.	immunoblot analysis using antibody specific to yeast ras-related protein yp20 revealed a 21kda protein associated with hela cell chromatin which immunologically cross-reacted with this ras-related protein. the 21kda protein can be rapidly released from the hela nuclei by micrococcal nuclease digestion. it seems to be associated with long mononucleosomes as determined by separation of chromatin subunits on sucrose gradients. we suggest that the 21kda hela protein may be the human homologue of yea ...	1990	2334423
transcriptional activation domain of the muscle-specific gene-regulatory protein myf5.	the human muscle determination factor myf5, like myod and other members of the family of skeletal muscle-specific regulatory proteins, contains a highly conserved putative helix-loop-helix domain. in myod this motif is required for the initiation of myogenesis in c3h mouse 10t1/2 fibroblasts and other non-muscle cells as well as for transcriptional activation of muscle genes. high affinity dna binding of myod to regulatory dna elements in muscle genes requires the formation of heterodimers with ...	1990	2385294
[circulating immune complexes and myasthenia gravis (analysis of 78 cases)].	for the sake of studying the circulating immune complexes (cic) of 78 patients with myasthenia gravis (mg), the agglutination assay of c3b sensitized with yeast cell (c3bsyca) was used. the anti human c3 elisa was also used for 21 out of the 78 patients. the positive results in the entire group of the patients studied were found to reach 82% and the igg and igm cic were detected also with anti-human c3 elisa. the titres of cic thus determined decreased as the clinical conditions improved. it was ...	1990	2390882
occurrence of s-(1,2-dicarboxyethyl)-cysteine at position 77 in mutant human lysozyme secreted by saccharomyces cerevisiae.	a mutant human lysozyme p110, in which val110 was replaced with pro, was secreted by saccharomyces cerevisiae; modification of the cysteine residue at position 77 was found in a purified mutant protein (p110-b) upon primary structure analysis. a peptide fragment containing 15 amino acid residues from thr70 to leu84 was obtained by proteolytic digestion of the protein and subsequently isolated by reverse-phase hplc. this fragment was analyzed by high-resolution fast-atom-bombardment (fab) mass sp ...	1990	2404763
generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes.	mammalian dna segments cloned as yeast artificial chromosomes (yacs) can be manipulated by dna-mediated transformation when placed in an appropriate yeast genetic background. a "fragmenting vector" has been developed that can introduce a yeast telomere and selectable marker into human-derived yacs at specific sites by means of homologous recombination, deleting all sequences distal to the recombination site. a powerful application of the method uses a human alu family repeat sequence to target r ...	1990	2406718
protein n-myristoylation in escherichia coli: reconstitution of a eukaryotic protein modification in bacteria.	protein n-myristoylation refers to the covalent attachment of a myristoyl group (c14:0), via amide linkage, to the nh2-terminal glycine residue of certain cellular and viral proteins. myristoyl-coa:protein n-myristoyltransferase (nmt) catalyzes this cotranslational modification. we have developed a system for studying the substrate requirements and biological effects of protein n-myristoylation as well as nmt structure-activity relationships. expression of the yeast nmt1 gene in escherichia coli ...	1990	2406721
antigenic conservation of primary structural regions of s-adenosylmethionine synthetase.	although the physical and kinetic properties of s-adenosylmethionine (adomet) synthetases from different sources are quite different, it appears that these enzymes have structurally or antigenically conserved regions as demonstrated by studies with adomet synthetase specific antibodies. polyclonal anti-human lymphocyte adomet synthetase crossreacted with enzyme from rat liver (beta isozyme), escherichia coli and yeast. in addition, polyclonal anti-e. coli enzyme and antibodies to synthetic pepti ...	1990	1698095
characterization of the human immunodeficiency virus type-1 reverse transcriptase enzyme produced in yeast.	the reverse transcriptase (rt) of human immunodeficiency virus type-1 (hiv-1) is comprised of two subunits of approximately 66kd and 51kd. we have defined the carboxyl terminus of the 51kd molecule using the 66kd rt and hiv-1 protease (pr) expressed in yeast. precise constructs encoding the 66kd and 51kd molecules were expressed individually, in yeast, at high levels. the purified recombinant subunits were shown to associate into heterodimers that retained both rt and rnase h activities. only th ...	1990	1698361
complementation of the mitotic activator, p80cdc25, by a human protein-tyrosine phosphatase.	the onset of m phase requires the activation of the pp34 protein kinase in all eukaryotes thus far examined. in schizosaccharomyces pombe, pp34 is phosphorylated on tyr15, and dephosphorylation of this residue regulates the initiation of mitosis. in this study, it is shown that dephosphorylation of tyr15 triggered activation of the pp34-cyclin complex from fission yeast, that a human protein-tyrosine phosphatase can catalyze this event both in vitro and in vivo, and that activation of fission ye ...	1990	1703321
integration of heterologous genes into the chromosome of saccharomyces cerevisiae using a delta sequence of yeast retrotransposon ty.	distribution of a delta (delta) sequence of the ty element on a chromosome of the yeast saccharomyces cerevisiae was analysed by pulsed-field gel electrophoresis. more than 100 copies of the delta sequence were nonrandomly distributed on the chromosome. using the delta sequence as a recombination site, mouse alpha-amylase and human beta-endorphin genes were introduced into the chromosomal dna. the integration occurred on a particular chromosome in each case and the copy number was estimated as t ...	1990	1369269
identification and characterization of an rna molecule that copurifies with rnase p activity from hela cells.	an rna molecule, 340 nucleotides in length and designated h1 rna, copurifies with rnase p activity from extracts of hela cells or isolated hela cell nuclei. when the genomic dna of various organisms is probed with h1 cdna in southern hybridization assays, only mammalian dna gives a positive signal. the gene coding for h1 rna in human cells is present in one to three copies per cell. the nucleotide sequence of h1 rna, which shows little homology to the known sequences of its analogs from prokaryo ...	1989	2470644
conserved pattern of antisense overlapping transcription in the homologous human ercc-1 and yeast rad10 dna repair gene regions.	we report that the genes for the homologous saccharomyces cerevisiae rad10 and human ercc-1 dna excision repair proteins harbor overlapping antisense transcription units in their 3' regions. since naturally occurring antisense transcription is rare in s. cerevisiae and humans (this is the first example in human cells), our findings indicate that antisense transcription in the ercc-1-rad10 gene regions represents an evolutionarily conserved feature.	1989	2471070
fine structure analysis of the chinese hamster as gene encoding asparagine synthetase.	overlapping cdnas for chinese hamster ovary (cho) asparagine synthetase (as) were isolated from a library prepared from an as-overproducing cell line. the sequence was determined and shown to contain an open reading frame encoding a protein of mr 64,300. the predicted amino acid sequence for the cho as enzyme was compared to that of the human as enzyme and found to be 95% homologous. a potential glutamine amide transfer domain, with sequence similarity to amidotransferases from bacteria and yeas ...	1989	2477309
proliferating cell nuclear antigen/cyclin in the ciliate euplotes eurystomus: localization in the replication band and in micronuclei.	human autoimmune sera specific for proliferating cell nuclear antigen (pcna)/cyclin (auxiliary protein for dna polymerase delta) demonstrated the presence of epitopes within the macro- and micronuclei of the hypotrichous ciliated protozoa euplotes eurystomus. tightly bound pcna/cyclin was localized at the site of dna synthesis in macronuclei, the rear zone of the replication band. starvation or heat shock, conditions that reduce macronuclear replication, resulted in a decrease of pcna/cyclin in ...	1989	2477376
conserved repetitive epitope recognized by cd4+ clones from a malaria-immunized volunteer.	t cell clones obtained from a human volunteer immunized with plasmodium falciparum sporozoites specifically recognized the native circumsporozoite (cs) antigen expressed on p. falciparum sporozoites, as well as bacteria- and yeast-derived recombinant falciparum cs proteins. the response of these cd4+ cd8- cells was species-specific, since the clones did not proliferate or secrete gamma interferon when challenged with sporozoites or recombinant cs proteins of other human, simian, or rodent malari ...	1989	2480642
characterization and expression of the human rhoh12 gene product.	the rho genes constitute an evolutionarily conserved family having significant homology to the ras oncogene family. these genes have been found in saccharomyces cerevisiae, drosophila melanogaster, rat, and human; their 21,000-dalton products show strong conservation of structure. in humans, three classes of rho cdna clones have been identified which differ by virtue of the presence of variable c-terminal domains: rhoh12, rhoh6, and rhoh9. the predicted 193 amino acids of human rhoh12 protein sh ...	1989	2501657
the 86-kilodalton antigen from schistosoma mansoni is a heat-shock protein homologous to yeast hsp-90.	we report the sequence of a cdna clone encoding an 86-kda polypeptide antigen (p86) from schistosoma mansoni. fusion proteins made in escherichia coli are recognized by human infection sera. the reading frame of this antigen is highly homologous to those of the large heat-shock proteins of saccharomyces cerevisiae (hsp90) and drosophila melanogaster (hsp83). mrna encoding p86 increases in response to heat shock of adult worms, as does hsp70. comparisons of the sequences of hsp70 and hsp83 homolo ...	1989	2509907
modulation of anti-candida activity of human alveolar macrophages by interferon-gamma or interleukin-1-alpha.	the fungicidal and bactericidal activities of human alveolar macrophages (am) and peripheral blood monocytes (pbm) from 18 healthy volunteers were evaluated. the results showed that am were able to phagocytize and kill candida albicans, pseudomonas aeruginosa, and staphylococcus aureus. however, killing of the bacteria was already complete in 2 h, whereas killing of candida required 4 to 6 h despite an early phagocytosis of yeast cells. the fungicidal activity of freshly collected am and pbm was ...	1989	2516451
recombinant soluble fc epsilon receptor ii (fc epsilon rii/cd23) has ige binding activity but no b cell growth promoting activity.	we have undertaken the production of recombinant soluble fc epsilon receptor ii (fc epsilon rii) as a secretory protein, but not as a cleavage product of membrane-bound receptor. several plasmid constructs containing soluble receptor sequence were prepared. only a chimeric gene containing the sequences encoding il-6 signal peptide and the soluble moiety of fc epsilon rii could be expressed in xenopus laevis oocytes and cho cells, resulting in the secretion of soluble fc epsilon rii. the recombin ...	1989	2523938
the association of the autoantigens of primary biliary cirrhosis with the mitochondrial h+-atpase--a reassessment.	the claimed association between the m2 autoantigens of primary biliary cirrhosis (pbc) and the mitochondrial h+-atpase has been re-examined in view of the recent reports that pbc autoantibodies react specifically with the lipoate acetyl transferases of 2-oxo acid dehydrogenases. study of f0f1-atpase purified from human and yeast mitochondria, and the comparison between immunoprecipitates obtained with antibodies against the h+-atpase beta subunit and anti-m2 antibodies of pbc, established that t ...	1989	2528956
increased plasma and erythrocyte selenium concentrations but decreased erythrocyte glutathione peroxidase activity after selenium supplementation in children with down syndrome.	an investigation was made of the activity of glutathione peroxidase (gsh-px) in erythrocytes and the levels of selenium in plasma and erythrocytes before, during and after selenium supplementation in children with down syndrome (ds). this subject is of interest since it has been suggested that selenium supplementation could enhance the gsh-px activity in erythrocytes, probably leading to improved protection against oxygen radicals, which might cause damage by lipid peroxidation, especially in th ...	1989	2532445
the application of molecular biology to the development of novel vaccines.	in summary, we have shown that yeast is the preferred host for the expression of recombinant-derived hepatitis b vaccines, and that a yeast expression system which is productive, stable and scaleable can be developed for each of the three hbv envelope proteins. the versatility of regulated and integrated yeast expression systems in the production of foreign polypeptides with biomedical utility also has been highlighted. we also have shown that careful attention to the development of recombinant ...	1989	2532858
[human epidermal growth factor gene is expressed in saccharomyces cerevisiae].	saccharomyces cerevisiae was transformed with plasmids containing the synthetic hegf gene under control of a alpha-factor promoter. the expressed protein was efficiently secreted into the medium broth and was shown to be a biologically active hegf.	1989	2534612
inhibition of leishmania donovani promastigote internalization into murine macrophages by chemically defined parasite glycoconjugate ligands.	leishmania donovani, the agent of human visceral leishmaniasis, is an intracellular parasite that must be recognized and internalized by host macrophages to complete its biological cycle. in a search for possible ligands for macrophage surface receptors, glycoconjugates were obtained from leishmania promastigotes by aqueous, phenol-aqueous, and alkaline extraction. a fucose-mannose glycoproteic ligand, a lipopeptidephosphoglycan, and a phosphate mannogalactan ligand were purified from promastigo ...	1989	2537257
mammalian camp-responsive element can activate transcription in yeast and binds a yeast factor(s) that resembles the mammalian transcription factor anf.	the human atf and ap1 transcription factors bind to highly related dna sequences. their consensus binding sites differ by a single nucleotide, but this single change is crucial in determining factor binding specificity. we have previously identified an ap1 (yap1) binding activity in yeast. in this report we identify a yeast atf (yatf) binding activity whose specificity can be distinguished from that of yap1 by the same crucial nucleotide that distinguishes binding of human atf and ap1. the atf b ...	1989	2538834
epstein-barr virus bzlf1 trans-activator specifically binds to a consensus ap-1 site and is related to c-fos.	two regions of the epstein-barr virus bzlf1 trans-activator protein have sequence similarity to the c-fos protein. part of the similarity corresponds to the region of c-fos which is similar to the dna binding domain of c-jun and gcn-4. the structure of the exon which contains this region in c-fos and bzlf1 is also highly conserved between the two genes. complete bzlf1 protein and a c terminal fragment were prepared either as purified fusion proteins or by in vitro translation from a bzlf1 cdna. ...	1989	2540954
the cdc2 kinase is a nuclear protein that is essential for mitosis in mammalian cells.	a homolog of the fission yeast cdc2-encoded protein kinase (p34) is a component of m phase promoting factor in xenopus oocytes. the homologous kinase in human hela cells is maximally active during mitosis, suggesting a mitotic role in mammalian somatic cells. this has been directly investigated by microinjection of anti-p34 antibodies into serum-stimulated rat fibroblasts. dna synthesis was unaffected but cell division was quantitatively blocked in injected cells. injection of antibodies against ...	1989	2541912
construction of yeast artificial chromosome libraries with large inserts using fractionation by pulsed-field gel electrophoresis.	a method for constructing yeast artificial chromosome (yac) libraries with large insert sizes is reported. high molecular weight human dna was partially digested with ecori and cloned in the vector pyac4. when unfractionated dna was used, the mean yac size was 120kb. fractionation by pulsed-field gel electrophoresis using a 'waltzer' apparatus to remove small dna fragments increased the mean yac size to congruent to 220kb or congruent to 370kb depending on the fractionation conditions. ligated d ...	1989	2542900
isolation of single-copy human genes from a library of yeast artificial chromosome clones.	a recently developed cloning system based on the propagation of large dna molecules as linear, artificial chromosomes in the yeast saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. two single-copy genes have now ...	1989	2544027
the ras oncogene--an important regulatory element in lower eucaryotic organisms.	the ras proto-oncogene in mammalian cells encodes a 21-kilodalton guanosine triphosphate (gtp)-binding protein. this gene is frequently activated in human cancer. as one approach toward understanding the mechanisms of cellular transformation by ras, the function of this gene in lower eucaryotic organisms has been studied. in the yeast saccharomyces cerevisiae, the ras gene products serve as essential function by regulating cyclic adenosine monophosphate metabolism. stimulation of adenylyl cyclas ...	1989	2547147
mapping of the active site tyrosine of eukaryotic dna topoisomerase i.	dna topoisomerase i from the yeasts saccharomyces cerevisiae and schizosaccharomyces pombe was overproduced using the cloned genes. extracts from cells overproducing dna topoisomerase i were prepared and incubated with 32p-labeled dna. alkali was used to trap the topoisomerase i-dna covalent intermediate. most of the dna was digested with nuclease, and the resultant 32p-labeled topoisomerase i was subjected to cleavage with cyanogen bromide or formic acid. from the molecular weights of the resul ...	1989	2547758
complementary dna encoding core protein ii of human mitochondrial cytochrome bc1 complex. substantial diversity in deduced primary structure from its yeast counterpart.	core protein ii of mitochondrial cytochrome bc1 complex is the second largest nuclear-encoded subunit that is essential for the assembly of the functional complex. we have isolated, for the first time, a cdna clone for a mammalian core protein ii. immunoscreening of a lambda gt11 rat liver cdna library resulted in isolation of a cdna encoding a partial sequence of rat mitochondrial core protein ii, and this cloned cdna was used as a probe for colony hybridization to screen a human fibroblast cdn ...	1989	2547763
isolation and characterization of a human telomere.	a method is described that allows cloning of human telomeres in s. cerevisiae by joining human telomeric restriction fragments to yeast artificial chromosome halves. the resulting chimeric yeast-human chromosomes propagate as true linear chromosomes, demonstrating that the human telomere structure is capable of functioning in yeast and suggesting that telomere functions are evolutionarily conserved between yeast and human. one cloned human telomere, yht1, contains 4 kb of human genomic dna seque ...	1989	2549507
temporary complementation of temperature-sensitive mutants of the cell cycle by transfection with a wild-type or a mutant cdna of adp/atp translocase.	a number of cell-cycle-specific temperature-sensitive (ts) mutants have been isolated from animal cells, especially syrian hamster cells. these ts mutants, like cell cycle ts mutants of yeast, can be complemented by specific genes, some of which have been molecularly cloned. we have isolated a cdna clone that complements tk-ts13 cells, but only temporarily. this clone, called b1, differs from a previously isolated clone (sekiguchi et al.: embo journal 7:1683-1687, 1988) that specifically complem ...	1989	2550485
expression of human dna topoisomerase i in yeast cells lacking yeast dna topoisomerase i: restoration of sensitivity of the cells to the antitumor drug camptothecin.	yeast saccharomyces cerevisiae strains that are permeable to the antitumor alkaloid camptothecin are killed by the drug if they express dna topoisomerase i, the cellular target of the drug (j. nitiss and j.c. wang, proc. natl. acad. sci. usa, 85: 7501-7505, 1988). we show that in a yeast strain permeable to camptothecin but lacking dna topoisomerase i, sensitivity to the drug was restored upon expression of human dna topoisomerase i from a plasmid-borne human complementary dna clone. when the hu ...	1989	2553253
molecular cloning of human uracil-dna glycosylase, a highly conserved dna repair enzyme.	uracil-dna glycosylase is the dna repair enzyme responsible for the removal of uracil from dna, and it is present in all organisms investigated. here we report on the cloning and sequencing of a cdna encoding the human uracil-dna glycosylase. the sequences of uracil-dna glycosylases from yeast, escherichia coli, herpes simplex virus type 1 and 2, and homologous genes from varicella-zoster and epstein-barr viruses are known. it is shown in this report that the predicted amino acid sequence of the ...	1989	2555154
phosphatidylinositol 3-kinase and its novel product, phosphatidylinositol 3-phosphate, are present in saccharomyces cerevisiae.	the metabolism of polyphosphoinositides has been shown to be an important factor in controlling the proliferation of saccharomyces cerevisiae. the monophosphate form of phosphatidylinositol has been assumed to be phosphatidylinositol 4-phosphate (pi-4-p). recent evidence from our laboratory has established that a phosphatidylinositol (pi) kinase, which phosphorylates the d-3 position of the inositol ring (pi 3-kinase), is associated with many activated protein-tyrosine kinases and may play an im ...	1989	2555343
fungispecificity of fluconazole against candida albicans.	candida albicans is an opportunistic pathogen of human mucosal surfaces. colonization of oral and vaginal mucosa by this yeast is antagonized by the resident normal bacterial population. however, antibacterial therapy can alter the normal flora to allow fungal cells to attach, grow and invade host tissues. we studied the antimicrobic activity of fluconazole against clinical isolates of oral and vaginal bacteria and candida albicans in vitro and in vivo by scanning and transmission electron micro ...	1989	2559330