Temperature-sensitive lethal mutant of era, a G protein in Escherichia coli.

T. Inada, K. Kawakami, S. M. Chen, H. E. Takiff, D. L. Court, Y. Nakamura

Research output: Contribution to journalArticlepeer-review

51 Citations (Scopus)

Abstract

The era gene of Escherichia coli encodes a GTP-binding protein which has similarities to elongation factor Tu and the Saccharomyces cerevisiae RAS protein. To investigate its function, mutations affecting era were isolated. A mini-Tn10 insertion, which truncated 22 amino acids from the COOH end of Era, did not affect cell growth. By using this mini-Tn10 insert as a coselectable marker, a temperature-sensitive lethal era mutant was isolated by localized mutagenesis using P1 phage transduction. A single-base G to A change was found at position 23, causing a tyrosine residue to be substituted for the cysteine residue at position 8 (era-770), in addition to the COOH-terminal mini-Tn10 disruption. Both alterations were necessary for the temperature-sensitive phenotype. Purified Era-770 mutant protein exhibited reduced binding to GTP compared with that of the wild-type Era protein.

Original languageEnglish
Pages (from-to)5017-5024
Number of pages8
JournalJournal of bacteriology
Volume171
Issue number9
DOIs
Publication statusPublished - 1989 Sep
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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