Cloning and sequencing of Schizosaccharomyces pombe car1 gene encoding arginase. Expression of the arginine anabolic and catabolic genes in response to arginine and related metabolites.
We report here the cloning and sequencing of the gene encoding arginase (car1) from Schizosaccharomyces pombe. Since no arginase-less strain exists in this organism, we cloned the gene by functional complementation of a car1 mutant strain from Saccharomyces cerevisiae. The S. pombe car1 gene encodes a 323 amino acids polypeptide sharing identity with arginases from different organisms. Measurements of arg3, arg11 and car1 mRNA under different growth conditions confirm the very weak repression by arginine of the two anabolic genes and show that the induction of arginase synthesis operates at a transcriptional level. The promoter of S. pombe car1 gene does not contain the 'arginine boxes' defined as the target of the ARGR-MCM1 proteins in the promoters of the arginine co-regulated genes in S. cerevisiae. The heterologous expression of S. pombe car1 gene in S. cerevisiae is independent of the ARGRII gene product (ArgRIIp/Arg81p). Determination of arginine, ornithine and citrulline intracellular concentrations shows the efficiency of the different controls operating in S. cerevisiae, and also indicates that in S. pombe enzyme compartmentation is not always sufficient to control the arginine metabolic flux.
Van Huffel
C
C
Institut de Recherches du CERIA, Université Libre de Bruxelles, Belgium.
Dubois
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E
Messenguy
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F
D000595:Amino Acid Sequence; D001119:Arginase / Q000235:genetics* / Q000378:metabolism; D001120:Arginine / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005800:Genes, Fungal / Q000235:genetics; D005816:Genetic Complementation Test; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009954:Ornithine Carbamoyltransferase / Q000235:genetics / Q000378:metabolism; D009953:Ornithine-Oxo-Acid Transaminase / Q000235:genetics / Q000378:metabolism; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000235:genetics / Q000378:metabolism; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D005656:Fungal Proteins; D008565:Membrane Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D001120:Arginine; D009954:Ornithine Carbamoyltransferase; D009953:Ornithine-Oxo-Acid Transaminase; D017851:Phosphotransferases (Carboxyl Group Acceptor); C018721:acetylglutamate kinase; D001119:Arginase; C490028:CAR1 protein, S cerevisiae; C490027:car1 protein, S pombe