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Isolation and characterization of ECT1 gene encoding CTP: phosphoethanolamine cytidylyltransferase of Saccharomyces cerevisiae
- Source :
- Journal of biochemistry. 120(5)
- Publication Year :
- 1996
-
Abstract
- 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 from 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.
Details
- ISSN :
- 0021924X
- Volume :
- 120
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Journal of biochemistry
- Accession number :
- edsair.pmid..........fe042cc61c24b3f5e469fbaf2079e72e