A cloned gene encoding phosphatidylserine decarboxylase complements the phosphatidylserine biosynthetic defect of a chinese hamster ovary cell mutant

Osamu Kuge, Masahiro Nishijima, Yuzuru Akamatsu

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Abstract

A phosphatidylseriae-auxotrophic mutant of cultured Chinese hamster ovary cells, PSA-3, manifests a defect in phosphatidylserine synthase I activity (Kuge, O., Nishijima, M., and Akamatsu, Y. (1986) J. Biol. Chem. 261, 5790-5794). We cloned a Chinese hamster gene, designated pssC, which was able to transform the PSA-3 cell line to a phosphatidylserine prototroph. The resultant transformant contained phosphatidylserine in normal amounts but remained defective in phosphatidylserine synthase I activity, indicating that pssC is a suppressor gene. Using the genomic fragment of pssC as a probe, a cDNA clone of pssC was isolated, and its nucleotide sequence was determined. A computer search through a protein data bank revealed that pssC had homology with the Escherichia coli psd gene encoding the proenzyme of phosphatidylserine decarboxylase at the amino acid level. Introduction of the cloned pssC gene into PSA-3 resulted in a 2-fold increase in phosphatidylserine decarboxylase activity. When the pssC cDNA was placed downstream of the yeast GAL1 promoter and introduced into yeast Saccharomyces cerevisiae cells, the phosphatidylserine decarboxylase activity increased in a galactose-dependent manner. These results indicate that pssC encodes phosphatidylserine decarboxylase. The mechanism by which pssC complements the defect of PSA-3 in phosphatidylserine biosynthesis is discussed.

Original languageEnglish
Pages (from-to)6370-6376
Number of pages7
JournalJournal of Biological Chemistry
Volume266
Issue number10
Publication statusPublished - 1991
Externally publishedYes

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Gene encoding
Phosphatidylserines
Cricetulus
Ovary
Cells
Yeast
Defects
Genes
Complementary DNA
Yeasts
Suppressor Genes
Enzyme Precursors
Biosynthesis
Galactose
Escherichia coli
Saccharomyces cerevisiae
Nucleotides
Clone Cells
Databases
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

A cloned gene encoding phosphatidylserine decarboxylase complements the phosphatidylserine biosynthetic defect of a chinese hamster ovary cell mutant. / Kuge, Osamu; Nishijima, Masahiro; Akamatsu, Yuzuru.

In: Journal of Biological Chemistry, Vol. 266, No. 10, 1991, p. 6370-6376.

Research output: Contribution to journalArticle

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