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Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase
The S. cerevisiae myristoyl-CoA:protein N-myristoyltransferase gene (NMT1) is essential for vegetative growth. NMT1 was found to be allelic with a previously described, but unmapped and unidentified mutation that causes myristic acid (C14:0) auxotrophy. The mutant (nmt1-181) is temperature sensitive...
Formato: | Texto |
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Lenguaje: | English |
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The Rockefeller University Press
1991
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2289034/ https://www.ncbi.nlm.nih.gov/pubmed/2045414 |
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collection | PubMed |
description | The S. cerevisiae myristoyl-CoA:protein N-myristoyltransferase gene (NMT1) is essential for vegetative growth. NMT1 was found to be allelic with a previously described, but unmapped and unidentified mutation that causes myristic acid (C14:0) auxotrophy. The mutant (nmt1-181) is temperature sensitive, but growth at the restrictive temperature (36 degrees C) is rescued with exogenous C14:0. Several analogues of myristate with single oxygen or sulfur for methylene group substitutions partially complement the phenotype, while others inhibit growth even at the permissive temperature (24 degrees C). Cerulenin, a fatty acid synthetase inhibitor, also prevents growth of the mutant at 24 degrees C. Complementation of growth at 36 degrees C by exogenous fatty acids is blocked by a mutation affecting the acyl:CoA synthetase gene. The nmt1-181 allele contains a single missense mutation of the 455 residue acyltransferase that results in a Gly451----Asp substitution. Analyses of several intragenic suppressors suggest that Gly451 is critically involved in NMT catalysis. In vitro kinetic studies with purified mutant enzyme revealed a 10-fold increase in the apparent Km for myristoyl-CoA at 36 degrees C, relative to wild-type, that contributes to an observed 200-fold reduction in catalytic efficiency. Together, the data indicate that nmt-181 represents a sensitive reporter of the myristoyl-CoA pools utilized by NMT. |
format | Text |
id | pubmed-2289034 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1991 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-22890342008-05-01 Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase J Cell Biol Articles The S. cerevisiae myristoyl-CoA:protein N-myristoyltransferase gene (NMT1) is essential for vegetative growth. NMT1 was found to be allelic with a previously described, but unmapped and unidentified mutation that causes myristic acid (C14:0) auxotrophy. The mutant (nmt1-181) is temperature sensitive, but growth at the restrictive temperature (36 degrees C) is rescued with exogenous C14:0. Several analogues of myristate with single oxygen or sulfur for methylene group substitutions partially complement the phenotype, while others inhibit growth even at the permissive temperature (24 degrees C). Cerulenin, a fatty acid synthetase inhibitor, also prevents growth of the mutant at 24 degrees C. Complementation of growth at 36 degrees C by exogenous fatty acids is blocked by a mutation affecting the acyl:CoA synthetase gene. The nmt1-181 allele contains a single missense mutation of the 455 residue acyltransferase that results in a Gly451----Asp substitution. Analyses of several intragenic suppressors suggest that Gly451 is critically involved in NMT catalysis. In vitro kinetic studies with purified mutant enzyme revealed a 10-fold increase in the apparent Km for myristoyl-CoA at 36 degrees C, relative to wild-type, that contributes to an observed 200-fold reduction in catalytic efficiency. Together, the data indicate that nmt-181 represents a sensitive reporter of the myristoyl-CoA pools utilized by NMT. The Rockefeller University Press 1991-06-02 /pmc/articles/PMC2289034/ /pubmed/2045414 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Articles Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title | Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title_full | Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title_fullStr | Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title_full_unstemmed | Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title_short | Myristic acid auxotrophy caused by mutation of S. cerevisiae myristoyl- CoA:protein N-myristoyltransferase |
title_sort | myristic acid auxotrophy caused by mutation of s. cerevisiae myristoyl- coa:protein n-myristoyltransferase |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2289034/ https://www.ncbi.nlm.nih.gov/pubmed/2045414 |