Publications

alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex dna sources.	current efforts to map the human genome are focused on individual chromosomes or smaller regions and frequently rely on the use of somatic cell hybrids. we report the application of the polymerase chain reaction to direct amplification of human dna from hybrid cells containing regions of the human genome in rodent cell backgrounds using primers directed to the human alu repeat element. we demonstrate alu-directed amplification of a fragment of the human hprt gene from both hybrid cell and cloned ...	1989	2771952
human platelet glycoprotein ix: an adhesive prototype of leucine-rich glycoproteins with flank-center-flank structures.	the glycoprotein (gp) ib-ix complex on the surface of human platelets functions as the von willebrand factor receptor and mediates von willebrand factor-dependent platelet adhesion to blood vessels. gpix is a relatively small (mr, 17,000) protein that may provide for membrane insertion and orientation of the larger component of the complex, gpib (mr, 165,000). using antibody screening, we cloned a cdna encoding gpix from a human erythroleukemia cell cdna library constructed in phage lambda gt11. ...	1989	2771955
v-cbl, an oncogene from a dual-recombinant murine retrovirus that induces early b-lineage lymphomas.	cas ns-1 is an acutely transforming murine retrovirus that induces pre-b and pro-b cell lymphomas. molecular cloning showed it was generated from the ecotropic cas-br-m virus by sequential recombinations with endogenous retroviral sequences and a cellular oncogene. the oncogene sequence shows no homology with known oncogenes but some similarity to the yeast transcriptional activator gcn4. a 100-kda gag-cbl fusion protein, with no detectable kinase activity, is responsible for the cellular transf ...	1989	2784003
arginase of agrobacterium ti plasmid c58. dna sequence, properties, and comparison with eucaryotic enzymes.	agrobacterium nopaline ti plasmids code for three enzymes of nopaline [n2-(1,3-dicarboxypropyl)-l-arginine] degradation: nopaline oxidase, arginase, and ornithine cyclodeaminase. we describe the dna sequence of the arginase gene, a comparison of the deduced protein sequence with eucaryotic arginases, and properties of the procaryotic enzyme. the results show that the agrobacterial arginase is related with arginases from yeast, rat liver, and human liver (28-33% identity). the ti plasmid enzyme r ...	1989	2806247
high levels of circulating neutralizing antibody in normal animals to recombinant mouse interferon-beta produced in yeast.	plasma from normal outbred swiss mice not previously given interferon (ifn) neutralized a glycosylated (high-mannose) recombinant murine ifn-beta made in yeast (rmuifn-beta y). the neutralization antibody titers were as high as 1:6,000 versus rmuifn-beta y, whereas the titers obtained with native murine ifn-beta (muifn-beta) were 100-fold less; a recombinant murine ifn-beta make in escherichia coli (rmuifn-beta ec) was not neutralized at 1:10 dilution of plasma. an elisa using rmuifn-beta y demo ...	1989	2809279
alpha-factor directed expression of the human epidermal growth factor in saccharomyces cerevisiae.	expression kinetics of the human epidermal growth factor (hegf) from the alpha-factor prepro region in a 2-mum based plasmid was studied in saccharomyces cerevisiae. production of hegf was highly medium de pendent as a chemically defined, nonenriched media had a significantly lower yield than did enriched media. also cells grown on yeast nitrogen base without amino acids with casamino acids degraded the hegf after cell growth as opposed to a yeast extract, peptone, and dextrose (yepd) medium, wh ...	1989	18588011
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
the molecular basis of genetic disease.	the pace of localization and characterization of genes affected in human genetic disorders is quickening. many important genes were localized or characterized recently: genes for in cystic fibrosis, nf-2, marfan's syndrome and xeroderma pigmentosum, to name a few. also, in the past 15 months, the cftr gene affected in cystic fibrosis has been isolated, the first disease gene to be isolated without use of previous cytogenetic clues, such as deletions or translocations in sporadic cases. other exa ...	1990	1367855
molecular analysis of the ets genes and their products.	organisms from human to drosophila have been found to contain cellular sequences and transcripts that are homologous to the ets region of the avian retrovirus, e26. ets-related sequences are present on at least two distinct functional loci in chickens and mammals, and have been designated ets-1 and ets-2. the e26 virus transduced sequences from the chicken ets-1 locus, which encompasses over 60 kb of dna. the ets genes characterized so far from sea urchin and drosophila are most closely related ...	1990	1964597
development of antibodies to unprotected glycosylation sites on recombinant human gm-csf.	in 4 out of 16 patients receiving recombinant human granulocyte macrophage colony stimulating factor (rhgm-csf) in phase i/ii studies antibodies developed to the recombinant protein. the antibodies react with sites on the native protein backbone which are normally protected by o-linked glycosylation but which are exposed in rhgm-csf produced in yeast and escherichia coli. antigenicity of recombinant human proteins due to non glycosylation may have relevance to the choice of host system for produ ...	1990	1968169
physical and genetic mapping of polymorphic loci in xq28 (dxs15, dxs52, and dxs134): analysis of a cosmid clone and a yeast artificial chromosome.	sequences corresponding to the xq28 loci dxs15, dxs52, dxs134, and dxs130 were shown to be present in a 140-kb yeast artificial chromosome (yac xy58, isolated by little et al.). this yac clone appears to contain a faithful copy of this genomic region, as shown by comparison with human dna and with a cosmid clone that contains probes st14c (part of the dxs52 sequences) and cpx67 (dxs134). cpx67 and st14c are contained in 11 kb and detect the same mspi rflp polymorphism. a comparison of the yac re ...	1990	1969226
molecular cloning, structure and expression of the yeast proliferating cell nuclear antigen gene.	the budding yeast saccharomyces cerevisiae is proving to be an useful and accurate model for eukaryotic dna replication. it contains both dna polymerase alpha (i) and delta (iii). recently, proliferating cell nuclear antigen (pcna), which in mammalian cells is an auxiliary subunit of dna polymerase delta and is essential for in vitro leading strand sv40 dna replication, was purified from yeast. we have now cloned the gene for yeast pcna (pol30). the gene codes for an essential protein of 29 kda, ...	1990	1970160
homologous sugar transport proteins in escherichia coli and their relatives in both prokaryotes and eukaryotes.	separate proteins for proton-linked transport of d-xylose, l-arabinose, d-galactose, l-rhamnose and l-fucose into escherichia coli are being studied. by cloning and sequencing the appropriate genes, the amino acid sequences of proteins for d-xylose/h+ symport (xyle), l-arabinose/h+ symport (arae), and part of the protein for d-galactose/h+ symport (galp) have been determined. these are homologous, with at least 28% identical amino acid residues conserved in the aligned sequences, although their ...	1990	1970645
nucleotide sequence of mouse hsp60 (chaperonin, groel homolog) cdna.	the cdna sequence of the 60 kda heat-shock protein from mouse 3t3 cells has been determined. the deduced amino acid sequence of mouse hsp60 protein differs from the corresponding proteins from chinese hamster and human cells in 7 and 13 residues, respectively, most of which are conservative replacements.	1990	1979012
furin is a subtilisin-like proprotein processing enzyme in higher eukaryotes.	the human fur gene encodes a protein, designated furin, the c-terminal half of which contains a transmembrane and a cysteine-rich receptor-like domain. the n-terminal half of furin exhibits striking primary amino acid sequence similarity to the catalytic domains of members of the subtilisin family of serine proteases. we here report characteristics of the furin protein and propose a three-dimensional model for its presumptive catalytic domain with characteristics, that predict furin to exhibit a ...	1990	2094803
enzymatic semisynthesis of human insulin: an update.	peptide bond formation can be enzymatically catalysed by the reverse reaction of proteases. application is seen in the industrial production of human insulin. human insulin derivative can be enzymatically prepared using either porcine insulin or the single chain b(1-29)-a(1-21) insulin precursor as the starting material. this is accomplished by either (1) digesting the starting material at lys29 with achromobacter lyticus protease i (ach) and then coupling with thr-x (x = blocking residue) (two- ...	1990	2096884
nucleotide sequence of citrus limon 26s rrna gene and secondary structure model of its rna.	the complete nucleotide sequence of citrus limon 26s rdna has been determined. the sequence has been aligned with large ribosomal rna (l-rrna) sequences of escherichia coli, saccharomyces cerevisiae and oryza sativa. nine extensive expansion segments in dicot 26s rrna relative to e. coli 23s rrna have been identified and compared with analogous segments of monocot, yeast, amphibian and human l-rrnas. a secondary structure model for lemon 26s rrna has been derived based on the refined model of e. ...	1990	2101688
distribution and scatter of yeast cell phagocytosis by human monocytes in an improved glass surface assay.	the assessment of yeast cell phagocytosis by glass-adherent monocytes was improved by ultrasonication of yeast cells prior to the experiments in order to prevent aggregation, restriction of measurements to completed engulfment regardless of the number of peptides ingested, and stopping of phagocytosis before counting by maintained cooling and the addition of edta. the modifications provided a reduced dispersion of individual values, increased engulfment of yeast cells, and decreased numbers of y ...	1990	2101806
characterization of a nonglycosylated single chain urinary plasminogen activator secreted from yeast.	using site-directed mutagenesis, we have changed the asparagine in human single-chain urinary plasminogen activator (u-pa) at position 302 to an alanine. this alteration removes the only known amino acid residue glycosylated in the protein. the single-chain u-pa containing an alanine residue at position 302 instead of asparagine (scu-pa(n302a] cdna gene was expressed in the yeast saccharomyces cerevisiae. secretion of the protein product into the culture broth was achieved by replacing the human ...	1990	2104831
the intracellular fate of histoplasma capsulatum in human macrophages is unaffected by recombinant human interferon-gamma.	human alveolar, peritoneal, and cultured macrophages were exposed in vitro to human recombinant interferon-gamma (rhuifn-gamma) and were tested for their ability to inhibit intracellular replication of yeast-phase histoplasma capsulatum. exposure at various concentrations, and for different time periods, failed to activate the macrophages to inhibit multiplication of intracellular yeast. macrophages were, however, activated by rhuifn-gamma as shown by their ability to inhibit intracellular repli ...	1990	2104917
doublet preference and gene evolution.	doublet preference analysis was carried out on coding and noncoding regions of escherichia coli, saccharomyces cerevisiae, and human mitochondrial and nuclear dna. the preference pattern in 1-2 and 2-3 doublets in e. coli and s. cerevisiae correlated with that in noncoding regions. the 3-1 doublet preference in e. coli genes with low optimal codon frequency and in s. cerevisiae genes also showed a correlation with each of their noncoding doublet preference. a mechanism to explain these double pr ...	1990	2107327
the 'effective number of codons' used in a gene.	a simple measure is presented that quantifies how far the codon usage of a gene departs from equal usage of synonymous codons. this measure of synonymous codon usage bias, the 'effective number of codons used in a gene', nc, can be easily calculated from codon usage data alone, and is independent of gene length and amino acid (aa) composition. nc can take values from 20, in the case of extreme bias where one codon is exclusively used for each aa, to 61 when the use of alternative synonymous codo ...	1990	2110097
cytosolic free calcium elevation mediates the phagosome-lysosome fusion during phagocytosis in human neutrophils.	cytosolic free calcium ([ca2+]i) and fusion of secondary granules with the phagosomal membrane (phagosome-lysosome fusion, p-l fusion) were assessed in single adherent human neutrophils during phagocytosis of c3bi-opsonized yeast particles. neutrophils were loaded with the fluorescent dye fura2/am and [ca2+]i was assessed by dual excitation microfluorimetry. discharge of lactoferrin, a secondary granule marker into the phagosome was verified by immunostaining using standard epifluorescence, conf ...	1990	2110568
isolation of cdna clones using yeast artificial chromosome probes.	the cloning of large dna fragments of hundreds of kilobases in yeast artificial chromosomes, has simplified the analysis of regions of the genome previously cloned by cosmid walking. the mapping of expressed sequences within cosmid contigs has relied on the association of genes with sequence motifs defined by rare-cutting endonucleases, and the identification of sequence conservation between species. we reasoned that if the contribution of repetitive sequences to filter hybridizations could be m ...	1990	2115671
effect of yeast lactase enzyme on "colic" in infants fed human milk.		1990	2116510
the molecular characterization of prp6 and prp9 yeast genes reveals a new cysteine/histidine motif common to several splicing factors.	prp6 and prp9 thermosensitive (ts) mutants are affected in pre-mrna splicing and transport from the nucleus to the cytoplasm. prp6 and prp9 wild-type alleles have been sequenced. dna sequence analysis reveals homologies in the 5' and 3' non-coding regions, suggesting a common regulation of gene expression. prp6 and prp9 genes encode a 899 amino acid and a 530 amino acid protein, respectively. the prp6 protein has repeated motifs that evoke helix-loop-helix structures. both prp6 and prp9 proteins ...	1990	2118103
role of the two activating domains of the oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the anti-oestrogen 4-hydroxytamoxifen.	various oestrogen responsive reporter genes and vectors expressing truncated or chimeric human oestrogen receptors (her) containing either of the two independent her transcriptional activation functions (taf-1 and taf-2) have been transfected into hela cells, chicken embryo fibroblast (cef) or yeast cells to investigate the agonistic activity of the anti-oestrogen 4-hydroxytamoxifen (oht). we demonstrate that the agonistic effect of oht on the whole her is due to the cell-type and promoter-conte ...	1990	2118104
cotranslational glycosylation of proteins in systems depleted of protein disulphide isomerase.	the role of protein disulphide isomerase (pdi) and other resident proteins of the endoplasmic reticulum (er) lumen in co- and post-translational modification of secretory proteins has been studied in experiments on translation in vitro. we have devised procedures for extracting the lumenal content proteins of dog pancreas microsomal vesicles by alkaline buffer, or detergent washing, and for reconstitution of the depleted membrane fraction. when microsomal membranes are depleted of content by was ...	1990	2120042
the catalytic domain of the neurofibromatosis type 1 gene product stimulates ras gtpase and complements ira mutants of s. cerevisiae.	sequencing of the neurofibromatosis gene (nf1) revealed a striking similarity among nf1, yeast ira proteins, and mammalian gap (gtpase-activating protein). using both genetic and biochemical assays, we demonstrate that this homology domain of the nf1 protein interacts with ras proteins. first, expression of this nf1 domain suppressed the heat shock-sensitive phenotype of yeast ira1 and ira2 mutants. second, this nf1 domain, after purification as a glutathione s-transferase (gst) fusion protein, ...	1990	2121369
gene-scrambling mutagenesis: generation and analysis of insertional mutations in the alginate regulatory region of pseudomonas aeruginosa.	a novel method for random mutagenesis of targeted chromosomal regions in pseudomona aeruginosa was developed. this method can be used with a cloned dna fragment of indefinite size that contains a putative gene of interest. cloned dna is digested to produce small fragments that are then randomly reassembled into long dna inserts by using cosmid vectors and lambda packaging reaction. this dna is then transferred into p. aeruginosa and forced into the chromosome via homologous recombination, produc ...	1990	2121708
molecular cloning and expression of a g25k cdna, the human homolog of the yeast cell cycle gene cdc42.	g25k is a low-molecular-mass gtp-binding protein with a broad distribution in mammalian tissues. a cdna clone was isolated by using oligonucleotides corresponding to the partial amino acid sequence of purified human g25k. the cdna encodes an 191-amino-acid polypeptide containing gtp-binding consensus sequences and a putative farnesylation site at the c terminus. the sequence exhibits 50 and 70% identities to the mammalian rho and rac proteins, respectively, and an 80% identity to the saccharomyc ...	1990	2122236
yeast artificial chromosomes: tools for mapping and analysis of complex genomes.	libraries of yeast artificial chromosomes (yacs) are representative of complex genomes, and typical yacs contain single fragments of dna that are up to a megabase or more in size, are stable during growth, and are faithful to genomic dna. such yacs may permit (1) the use of complete gene units for a number of research and medical purposes; and (2) the assembly of chromosome-sized contigs in a single map that unifies genetic and physical data.	1990	2122561
performance characteristics of a novel immunoassay based on hybrid ty virus-like particles (ty-vlps): rapid differentiation between hiv-1 and hiv-2 infection.	recombinant antigens containing all or parts of the hiv-1 proteins p24, nef and gp41 and hiv-2 gp36 have been purified and used to develop a rapid immunoassay to detect and differentiate between hiv-1 and hiv-2 antibodies in a single test. the antigens were produced as particulate fusion proteins by exploiting the ability of a protein encoded by the yeast retrotransposon ty to assemble into virus-like particles (ty-vlps). hybrid hiv: ty-vlps carrying each of the antigens were applied to nitrocel ...	1990	2124493
methylation of intact chromosomes by bacterial methylases in agarose plugs suitable for pulsed-field electrophoresis. methylation of intact chromosomes in agarose by methylases.	conditions were determined for the methylation of intact yeast chromosomes by ecori, hhai, and mspi bacterial methylases using an endonuclease protection assay while the chromosomes were embedded in agarose plugs suitable for transverse-field electrophoresis. parameters were also established for the methylation of human chromosomes by ecori methylase. methylation of embedded chromosomes by ecori methylase required prewashes with edta. ecori, hhai, and mspi methylases showed optimal activity when ...	1990	2128170
phototumorigenesis studies of 5-methoxypsoralen in bergamot oil: evaluation and modification of risk of human use in an albino mouse skin model.	the skin of the female hairless albino mouse (skh 1) was used to study the enhancement of solar simulated radiation (ssr) tumorigenesis by 5-methoxypsoralen (5-mop) in model perfumes that contain bergamot oil. this work was done in association with yeast mutagenicity studies and human skin phototoxicity studies. analyses of time-to-onset of tumour observation with 5-mop at 0, 5, 15 and 50 ppm show a highly significant 5-mop dose effect and the data indicate that 5-mop has phototumorigenic potent ...	1990	2128326
reversion and rereversion in ras-transformed cells.		1990	2128917
translational regulation of the lymphocyte-specific protein tyrosine kinase p56lck.	the lck gene encodes a lymphocyte-specific membrane-associated protein tyrosine kinase that is implicated in signal transduction processes that regulate t-cell growth. previous studies demonstrate that lck transcripts, unlike most other vertebrate mrnas, contain 5' proximal aug sequences upstream relative to the major open reading frame (5' augs) that serve as satisfactory targets for recognition by 40s ribosomal subunits. there exist two distinct lck transcripts, generated through alternative p ...	1990	2133653
immobilization of galactosyltransferase and continuous galactosylation of glycoproteins in a reactor.	we immobilized human milk galactosyltransferase covalently to cnbr- and tresylchloride-activated sepharose. the enzyme was also immobilized non-covalently to concanavalin a-sepharose and to monoclonal anti-galactosyltransferase antibodies which were bound via their fc-fragment to protein g-sepharose. with the covalent methods, up to 72% of the enzyme could be bound to the carrier, but more than 90% of the specific activity was lost. in contrast, non-covalent immobilization yielded only about 50% ...	1990	2136355
unexpected flexibility in an evolutionarily conserved protein-rna interaction: genetic analysis of the sm binding site.	human autoantibodies of the sm specificity recognize a conserved set of proteins found in the u class small nuclear ribonucleoproteins (u snrnps), key trans-acting factors involved in the splicing of mrna precursors. the sm protein binding site in u snrnas is unusual because of its single-stranded nature and its simple sequence motif (au5-6gpu). here we use genetics to probe this specific protein-rna interaction by saturation mutagenesis of the sm binding site of the saccharomyces cerevisiae u5 ...	1990	2142451
presence of a potent transcription activating sequence in the p53 protein.	the p53 gene is frequently mutated in a wide variety of human cancers. however, the role of the wild-type p53 gene in growth control is not known. hybrid proteins that contain the dna binding domain of yeast gal4 and portions of p53 have been used to show that the p53 protein contains a transcription-activating sequence that functions in both yeast and mammalian cells. the nh2-terminal 73 residues of p53 activated transcription in mammalian cells as efficiently as the herpes virus protein vp16, ...	1990	2144363
structure, exon pattern, and chromosome mapping of the gene for cytosolic copper-zinc superoxide dismutase (sod-1) from neurospora crassa.	a 4.8-kilobase bamhi-hindiii fragment encoding the entire neurospora crassa cuzn superoxide dismutase gene (herein designated sod-1) was isolated from a genomic library using two 60-base deoxyoligonucleotide probes corresponding to the published n. crassa amino acid sequence. the nucleotide sequence of the gene encodes an amino acid sequence matching the published protein sequence at 152 of 153 positions. codon preference shows an unusually strong bias such that only 32 of the possible 61 codons ...	1990	2146266
evidence that specific high-mannose oligosaccharides can directly inhibit antigen-driven t-cell responses.	uromodulin is an 85 kd immunosuppressive glycoprotein originally isolated from human pregnancy urine. it is unique in that most of its biologic activity can be attributed to attached oligosaccharides. purified immunomodulatory oligosaccharides from uromodulin have been structurally characterized using 1h-nmr spectroscopy and shown to be man6-7glcnac2(m6,m7). based on these observations, we isolated high-mannose n-type oligosaccharides and glycopeptides from ovalbumin, soybean agglutinin, and yea ...	1990	2146352
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