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. 1989 Sep;171(9):5017–5024. doi: 10.1128/jb.171.9.5017-5024.1989

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

T Inada 1, K Kawakami 1, S M Chen 1, H E Takiff 1, D L Court 1, Y Nakamura 1
PMCID: PMC210312  PMID: 2527846

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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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