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Four Genes Responsible for a Position Effect on Expression from HML and HMR in Saccharomyces cerevisiae 总被引:64,自引:12,他引:52 下载免费PDF全文
Mating type interconversion in Saccharomyces cerevisiae occurs by transposition of copies of the a or alpha mating type cassettes from inactive loci, HML and HMR, to an active locus, MAT. The lack of expression of the a and alpha genes at the silent loci results from repression by trans-acting regulators encoded by SIR (Silent Information Regulator) genes. In this paper we present evidence for the existence of four SIR genes. Inactivation of any of these genes leads to expression of cassettes at both HML and HMR. Unusual complementation properties are observed for a number of sir mutations. Specifically, some recessive mutations in different genes fail to complement. The correspondence between SIR1, SIR2, SIR3, SIR4 and other genes with similar roles (MAR, CMT, STE8 and STE9) is presented. 相似文献
5.
R J Wanders P G Barth C W van Roermund R Ofman R Wolterman R B Schutgens J M Tager H van den Bosch P A Bolhuis 《Experimental cell research》1987,170(1):147-152
In the present study we investigated peroxisomal functions in cultured human muscle cells from control subjects and from a patient with the Zellweger syndrome, a genetic disease characterized by the absence of morphologically distinguishable peroxisomes in liver and kidney. In homogenates of cultured muscle cells from control subjects, catalase is contained within subcellular particles, acyl-CoA:dihydroxyacetonephosphate acyltransferase activity is present and palmitoyl-CoA can be oxidized by a peroxisomal beta-oxidative pathway; these findings are indicative of the presence of peroxisomes in the cells. In homogenates of cultured muscle cells from the patient with the Zellweger syndrome, acyl-CoA:dihydroxyacetonephosphate acyltransferase activity was deficient, peroxisomal beta-oxidation of palmitoyl-CoA was impaired and catalase was not particle-bound. These findings indicate that functional peroxisomes are absent in muscle from patients with the Zellweger syndrome. We conclude that cultured human muscle cells can be used as a model system to study peroxisomal functions in muscle and the consequences for this tissue of a generalized dysfunction of peroxisomes. 相似文献
6.
STE16, a New Gene Required for Pheromone Production by a Cells of Saccharomyces cerevisiae 总被引:9,自引:1,他引:8 下载免费PDF全文
Genes required for mating by a and alpha cells of Saccharomyces cerevisiae (STE, "sterile," genes) encode products such as peptide pheromones, pheromone receptors, and proteins responsible for pheromone processing. a-specific STE genes are those required for mating by a cells but not by alpha cells. To identify new a-specific STE genes, we have employed a novel strategy that enabled us to determine if a ste mutant defective in mating as a is also defective in mating as alpha without the need to do crosses. This technique involved a strain (K12-14b) of genotype mata1 HML alpha HMR alpha sir3ts, which mates as a at 25 degrees and as alpha at 34 degrees. We screened over 40,000 mutagenized colonies derived from K12-14b and obtained 28 a-specific ste mutants. These strains contained mutations in three known a-specific genes--STE2, STE6 and STE14--and in a new gene, STE16. ste16 mutants are defective in the production of the pheromone, a-factor, and exhibit slow growth. Based on the distribution of a-specific ste mutants described here, we infer that we have identified most if not all nonessential genes that can give rise to a-specific mating defects. 相似文献
7.
Ira D. Bhattacharya M. F. Picciano John A. Milner 《Biological trace element research》1988,18(1):59-70
Human milk glutathione peroxidase (GPx) was purified 4500-fold using acetone precipitation and purification by repetitive
ion-exchange and gel filtration chromatography with an overall yield of 34%. Homogeneity was established by gel electrophoresis.
Using gel filtration, the molecular weight (mol wt) of the enzyme was estimated to be 92 kdalton (kD). The monomeric molecular
weight was estimated to b 23 kD from polyacrylamide gel electrophoresis, indicating that the native enzyme consists of four
identical subunits. The molecular weight of each subunit was supported by amino acid analysis. Selenium (Se) content of the
purified enzyme was 0.31%, in a stoichiometry of 3.7 g-atoms/mol. Data from these studies reveal that GPx provided approximately
22% of total milk Se, but only 0.025% of the total protein. 相似文献
8.
J M Aerts J Heikoop S van Weely W E Donker-Koopman J A Barranger J M Tager A W Schram 《Experimental cell research》1988,177(2):391-398
We have characterized glucocerebrosidase in various cell types of peripheral blood of control subjects and in cultured human blastoid cells. The intracellular level of glucocerebrosidase in cultured blastoid cells (10-30 nmol substrate hydrolyzed/h.mg protein) resembles closely values observed for leukocyte cell types and various tissues and is significantly lower than that observed in cultured fibroblasts (150-500 nmol substrate hydrolyzed/h.mg protein). Glucocerebrosidase is extracted from leukocyte cell types and cultured blastoid cells almost exclusively in a monomeric, nonactivated form with enzymatic properties identical to those of the tissue enzyme. In contrast, extracts of platelets are rich in an aggregated, activated form of the enzyme. Glucocerebrosidase in blood cells and cultured blastoid cells is heterogeneous with respect to Mr and pI due to a heterogeneous oligosaccharide composition of the enzyme. The different forms seen represent intermediates in the biosynthesis and maturation of the enzyme. Blastoid cells should thus be an attractive model system for studying the natural history of glucocerebrosidase in a cell type related to those cells involved in the pathology of Gaucher disease. 相似文献
9.
J M Aerts W E Donker-Koopman M Koot G J Murray J A Barranger J M Tager A W Schram 《Biochimica et biophysica acta》1986,863(1):63-70
Human urine contains a soluble form of glucocerebrosidase, an enzyme associated with the lysosomal membrane in cells and tissues. Urinary glucocerebrosidase is identical to the enzyme extracted from tissues with respect to the following parameters: Km for natural and artificial substrates, inhibition by conduritol B-epoxide, and stimulation by taurocholate. The enzyme is greater than 90% precipitable by polyclonal anti-(placental glucocerebrosidase) antiserum. Upon isoelectric focussing of urinary glucocerebrosidase multiple peaks of activity were observed. Partial deglycosylation (removal of sialic acid, N-acetylglucosamine and galactose) of the urinary enzyme increased the isoelectric point to a value identical to that of the main form found after partial deglycosylation of the placental enzyme. Upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate followed by immunoblotting, the immunopurified urinary enzyme shows the same molecular mass forms as the enzyme immunopurified from brain and kidney. In placenta the apparent molecular mass is somewhat higher but upon removal of sialic acid, N-acetylglucosamine and galactose the urinary and the placental enzyme show identical molecular masses of 57 kDa. We conclude that the enzymes extracted from urine and tissue are identical and that differences in apparent molecular mass and isoelectric point are probably due to heterogeneity in the oligosaccharide moieties of the molecules. 相似文献
10.
Acyl-CoA:dihydroxyacetone phosphate acyltransferase in human skin fibroblasts: study of its properties using a new assay method 总被引:2,自引:0,他引:2
R B Schutgens G J Romeyn R Ofman H van den Bosch J M Tager R J Wanders 《Biochimica et biophysica acta》1986,879(3):286-291
In relation to the finding that human skin fibroblasts are capable of de novo either phospholipid biosynthesis, we have studied the properties of acyl-CoA:dihydroxyacetone phosphate acyltransferase in fibroblast homogenates using a new assay method. The results indicate that the acylation of dihydroxyacetone phosphate shows an optimum at pH 5.5 with a broad shoulder of activity up to pH 6.4 and a decline in activity up to pH 8.2. At pH 5.5 the acyltransferase accepts dihydroxyacetone phosphate, but not glycerol 3-phosphate as a substrate. Furthermore, the transferase activity was found to be membrane-bound and inactivated by Triton X-100 at concentrations above 0.025% (w/v). Similar properties have been described for the enzyme as present in rat-liver and guinea-pig liver peroxisomes. These data, together with the finding that acyl-CoA:dihydroxyacetone phosphate acyltransferase is deficient in cultured skin fibroblasts from patients without peroxisomes (Zellweger syndrome), suggest that in cultured skin fibroblasts the enzyme is primarily located in peroxisomes. 相似文献