Mutations of human myristoyl-CoA:protein N-myristoyltransferase cause temperature-sensitive myristic acid auxotrophy in Saccharomyces cerevisiae

Robert J. Duronio, Steven I. Reed, Jeffrey I. Gordon

Research output: Contribution to journalArticle

110 Scopus citations

Abstract

We have isolated cDNAs encoding human myristoyl-CoA:protein N-myristoyltransferase (NMT, EC 2.3.1.97) by complementing the nmt1-181 mutation of Saccharomyces cerevisiae, which causes temperature-sensitive myristic acid auxotrophy. Human NMT is derived from a single-copy gene, contains 416 amino acids, is 44% identical to S. cerevisiae NMT (yeast NMT), and can complement the lethal phenotype of an nmt1 null mutation. Human and yeast NMTs have overlapping yet distinct protein substrate specificities as judged by a coexpression system that reconstitutes protein N-myristoylation in Escherichia coli. Both enzymes contain a glycine five residues from the C terminus. Gly → Asp or Lys mutagenesis in these orthologous NMTs produces marked reductions in their activities in E. coli as well as temperature-sensitive myristic acid auxotrophy in S. cerevisiae. These results indicate highly con- served structure-function relationships in vivo and underscore the usefulness of these functional assays for identifying factors that regulate protein N-myristoylation in mammalian systems.

Original languageEnglish
Pages (from-to)4129-4133
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume89
Issue number9
DOIs
StatePublished - Jan 1 1992

Keywords

  • Complementation cloning
  • Drug design
  • Enzyme structure-function
  • Fatty acid metabolism
  • Protein N-myristoylation

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