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PMID: 18252723 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

YLR099C (ICT1) encodes a soluble Acyl-CoA-dependent lysophosphatidic acid acyltransferase responsible for enhanced phospholipid synthesis on organic solvent stress in Saccharomyces cerevisiae.

The Journal of biological chemistry ·Vol. 283 ·No. 15 ·2008-04-11 ·Pages 9768-75

Ghosh AK, Ramakrishnan G, Rajasekharan R

Abstract

One of the major determinants of organic solvent tolerance is the increase in membrane phospholipids. Here we report for the first time that an increase in the synthesis of phosphatidic acid is responsible for enhanced phospholipid synthesis that confers tolerance to the organic solvent in Saccharomyces cerevisiae. This increase in phosphatidic acid formation is because of the induction of Ict1p, a soluble oleoyl-CoA:lysophosphatidic acid acyltransferase. YLR099C (ICT1) was reported to be maximally expressed during solvent tolerance (Miura, S., Zou, W., Ueda, M., and Tanaka, A. (2000) Appl. Environ. Microbiol. 66, 4883-4889); however, its physiological significance was not understood. In silico analysis revealed the absence of any transmembrane domain in Ict1p. Domain analysis showed that it has a hydrolase/acyltransferase domain with a distinct lipid-binding motif and a lysophospholipase domain. Analysis of ict1Delta strain showed a drastic reduction in phosphatidic acid suggesting the role of Ict1p in phosphatidic acid biosynthesis. Overexpression of Ict1p in S. cerevisiae showed an increase in phosphatidic acid and other phospholipids on organic solvent exposure. To understand the biochemical function of Ict1p, the gene was cloned and expressed in Escherichia coli. The purified recombinant enzyme was found to specifically acylate lysophosphatidic acid. Specific activity of Ict1p was found to be higher for oleoyl-CoA as compared with palmitoyl- and stearoyl-CoAs. This study provides a mechanism for organic solvent tolerance from the point of membrane dynamics in S. cerevisiae.

MeSH Terms
Acyl Coenzyme A/genetics,metabolism Acyltransferases/biosynthesis,genetics Enzyme Induction/drug effects Gene Expression Regulation, Enzymologic/drug effects,physiology Gene Expression Regulation, Fungal/drug effects,physiology Phosphatidic Acids/biosynthesis,genetics Protein Structure, Tertiary/physiology Recombinant Proteins/genetics,metabolism Saccharomyces cerevisiae/enzymology,genetics Solvents/pharmacology
Chemicals
Acyl Coenzyme A Phosphatidic Acids Recombinant Proteins Solvents Acyltransferases 2-acylglycerophosphate acyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ghosh Ananda K
Department of Biochemistry, Indian Institute of Science, Bangalore 560012, India.
Ramakrishnan Geetha
Rajasekharan Ram
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M708418200
Published
2008-04-11
Epub
2008-00-04
Pages
9768-75
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2442289
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M708418200 PMC Full Text View journal details
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