Isolation and characterization of ECT1 gene encoding CTP: Phosphoethanolamine cytidylyltransferase of Saccharomyces cerevisiae

作者：MinSeok R; Kawamata Y; Nakamura H; Ohta A; Takagi M

来源：Journal of Biochemistry, 1996, 120(5): 1040-1047.

• CDP-ethanolamine
• Kennedy pathway
• phosphatidylethanolamine
• phosphoethanolamine cytidylyltransferase
• Saccharomyces cerevisiae

Saccharomyces cerevisiae mutants that were unable to utilize extracellular ethanolamine for phosphatidylethanolamine synthesis were isolated, Two of them carried recessive chromosomal mutations in a same gene and were defective in CTP: phosphoethanolamine cytidylyltransferase (ECT) activity in vitro (Ect(-)). In an Ect(-) mutant that also carried the cho1 mutation, phosphatidylethanolamine accounted for less than 2% of total phospholipids, suggesting the importance of ECT in phosphatidylethanolamine synthesis. By screening a genomic library on a low copy number vector, three complementary clones of different size were isolated, A 2.8-kb common DNA region carried an open reading frame (ORF) of 969 bp in length, of which a truncated form failed to complement the Ect(-) mutation, This ORF was identical to the previously isolated MUQ1 gene of unknown function, Its deduced amino acid sequence had significant similarity to CTP:phosphocholine cytidylyl-transferases of yeast and rat, The entire ORF, when combined with the glutathione S-transferase gene and expressed in Escherichia coli, exhibited ECT activity, These results indicate that the cloned gene encodes a catalytic subunit of ECT of S. cerevisiae.

• 出版日期1996-11