Analysis of the mutagenic properties of the UmuDC, MucAB and RumAB proteins, using a site-specific abasic lesion (original) (raw)

Abstract

The_mucAB_ and_rumAB_ loci have been shown to promote mutagenesis to a greater extent than the structurally and functionally homologous_Escherichia coli umuDC_ operon. We have analyzed the basis of this enhanced mutagenesis by comparing the influence of these operons, relative to_umuDC_, on the mutagenic properties of each of two abasic sites, specifically located in a single-stranded vector. Experiments with these vectors are useful analytical tools because they provide independent estimates of the efficiency of translesion synthesis and of the relative frequencies of each type of nucleotide insertion or other kind of mutagenic event. The_umuDC, mucAB_, and_rumAB_ genes were expressed from their natural_LexA_-regulated promoter on low-copy-number plasmids in isogenic strains carrying a_umuDC_ deletion. In addition, plasmids expressing the UmuD'C, MucA'B, or RumA'B proteins were also used. Compared to_umuDC_, the chief effect of_mucAB_ was to increase the efficiency of translesion synthesis past the abasic site. The enhanced capacity of_mucAB_ for translesion synthesis depended about equally on an inherently greater capacity to promote this process and on a greater susceptibility of the MucA protein to proteolytic processing. The RumA protein also appeared to be more susceptible to proteolytic processing, but the inherent capacity of the_Rum_ products for translesion synthesis was no greater than that of_UmuDC_. dAMP was inserted opposite one of the two abasic sites studied at a somewhat greater frequency in strains expressing_rum_ (82%) compared to those expressing_umu_ (72%), which might result in higher mutation frequencies in_rumAB_ than in_umuDC_ strains.

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Authors and Affiliations

1. Department of Biophysics, University of Rochester Medical Center, 14642, Rochester, NY, USA
   C. W. Lawrence & A. Borden
2. Section on DNA Replication, Repair and Mutagenesis, National Institute of Child Health and Human Development, National Institutes of Health, 20892, Bethesda, MD, USA
   R. Woodgate

Authors

1. C. W. Lawrence
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2. A. Borden
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3. R. Woodgate
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Communicated by B. J. Kilbey

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Lawrence, C.W., Borden, A. & Woodgate, R. Analysis of the mutagenic properties of the UmuDC, MucAB and RumAB proteins, using a site-specific abasic lesion.Molec. Gen. Genet. 251, 493–498 (1996). https://doi.org/10.1007/BF02172378

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• Received: 22 November 1995
• Accepted: 27 November 1995
• Issue Date: June 1996
• DOI: https://doi.org/10.1007/BF02172378

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