Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1989 Sep;86(17):6577-81.
doi: 10.1073/pnas.86.17.6577.

Mutations in the Escherichia coli UvrB ATPase motif compromise excision repair capacity

T W Seeley  1 , L Grossman
Affiliations

Mutations in the Escherichia coli UvrB ATPase motif compromise excision repair capacity

T W Seeley et al. Proc Natl Acad Sci U S A. 1989 Sep.

Abstract

The Escherichia coli UvrB protein possesses an amino acid sequence motif common to many ATPases. The role of this motif in UvrB has been investigated by site-directed mutagenesis. Three UvrB mutants, with amino acid replacements at lysine-45, failed to confer UV resistance when tested in the UV-sensitive strain N364 (delta uvrB), while five other mutants constructed near this region of UvrB confer wild-type levels of UV resistance. Because even the conservative substitution of arginine for lysine-45 in UvrB results in failure to confer UV resistance, we believe we have identified an amino acid side chain in UvrB essential to nucleotide excision repair in E. coli. The properties of two purified mutant UvrB proteins, lysine-45 to alanine (K45A) and asparagine-51 to alanine (N51A), were analyzed in vitro. While the K45A mutant is fully defective in incision of UV-irradiated DNA, K45A is capable of interaction with UvrA in forming an ATP-dependent nucleoprotein complex. The K45A mutant, however, fails to activate the characteristic increase in ATPase activity observed with the wild-type UvrB in the presence of UvrA and DNA. From these results we conclude that there is a second nucleotide-dependent step in incision following initial complex formation, which is defective in the K45A mutant. This experimental approach may prove of general applicability in the study of function and mechanism of other ATPase motif proteins.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Eur J Biochem. 1974 Mar 15;43(1):131-44 - PubMed
    1. J Biol Chem. 1986 Apr 15;261(11):4895-901 - PubMed
    1. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1326-30 - PubMed
    1. Nature. 1983 Jan 20;301(5897):262-4 - PubMed
    1. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6157-61 - PubMed

Publication types

LinkOut - more resources