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Journal of Bacteriology, September 1998, p. 4658-4666, Vol. 180, No. 17
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Novel Escherichia coli umuD' Mutants: Structure-Function Insights into SOS Mutagenesis

Mary McLenigan,¹ Thomas S. Peat,^2, Ekaterina G. Frank,¹ John P. McDonald,¹ Martín Gonzalez,¹ Arthur S. Levine,¹ Wayne A. Hendrickson,^2,3 and Roger Woodgate^1,*

Section on DNA Replication, Repair and Mutagenesis, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892¹ and Department of Biochemistry and Molecular Biophysics² and Howard Hughes Medical Institute,³ Columbia University, New York, New York 10032

Received 20 March 1998/Accepted 20 June 1998

Although it has been 10 years since the discovery that the Escherichia coli UmuD protein undergoes a RecA-mediated cleavage reaction to generate mutagenically active UmuD', the function of UmuD' has yet to be determined. In an attempt to elucidate the role of UmuD' in SOS mutagenesis, we have utilized a colorimetric papillation assay to screen for mutants of a hydroxylamine-treated, low-copy-number umuD' plasmid that are unable to promote SOS-dependent spontaneous mutagenesis. Using such an approach, we have identified 14 independent umuD' mutants. Analysis of these mutants revealed that two resulted from promoter changes which reduced the expression of wild-type UmuD', three were nonsense mutations that resulted in a truncated UmuD' protein, and the remaining nine were missense alterations. In addition to the hydroxylamine-generated mutants, we have subcloned the mutations found in three chromosomal umuD1, umuD44, and umuD77 alleles into umuD'. All 17 umuD' mutants resulted in lower levels of SOS-dependent spontaneous mutagenesis but varied in the extent to which they promoted methyl methanesulfonate-induced mutagenesis. We have attempted to correlate these phenotypes with the potential effect of each mutation on the recently described structure of UmuD'.

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^* Corresponding author. Mailing address: Building 6, Room 1A13, NICHD, NIH, 9000 Rockville Pike, Bethesda, MD 20892-2725. Phone: (301) 496-6175. Fax: (301) 594-1135. E-mail: woodgate{at}helix.nih.gov.

Present address: Los Alamos National Laboratory, Los Alamos, NM 87545.

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Journal of Bacteriology, September 1998, p. 4658-4666, Vol. 180, No. 17
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

This article has been cited by other articles:

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• Ohta, T., Sutton, M. D., Guzzo, A., Cole, S., Ferentz, A. E., Walker, G. C. (1999). Mutations Affecting the Ability of the Escherichia coli UmuD' Protein To Participate in SOS Mutagenesis. J. Bacteriol. 181: 177-185 [Abstract] [Full Text]