Host cell RecA activates a mobile element-encoded mutagenic DNA polymerase

Debika Ojha; Malgorzata M Jaszczur; Adhirath Sikand; John P McDonald; Andrew Robinson; Antoine M van Oijen; Chi H Mak; Fabien Pinaud; Michael M Cox; Roger Woodgate; Myron F Goodman

doi:10.1093/nar/gkac515

Host cell RecA activates a mobile element-encoded mutagenic DNA polymerase

Nucleic Acids Res. 2022 Jul 8;50(12):6854-6869. doi: 10.1093/nar/gkac515.

Authors

Debika Ojha¹, Malgorzata M Jaszczur¹, Adhirath Sikand², John P McDonald³, Andrew Robinson^{4

5}, Antoine M van Oijen^{4

5}, Chi H Mak^{1

2

6}, Fabien Pinaud^{1

2

7}, Michael M Cox⁸, Roger Woodgate³, Myron F Goodman^{1

2}

Affiliations

¹ Department of Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA.
² Department of Chemistry, University of Southern California, Los Angeles, Los Angeles, CA 90089, USA.
³ Laboratory of Genomic Integrity, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
⁴ Molecular Horizons Institute and School of Chemistry and Biomolecular Science, University of Wollongong, Wollongong, NSW 2522, Australia.
⁵ Illawarra Health and Medical Research Institute, Wollongong, NSW 2522, Australia.
⁶ Center of Applied Mathematical Sciences, University of Southern California, Los Angeles, CA 90089, USA.
⁷ Department of Physics and Astronomy, University of Southern California, Los Angeles, CA 90089, USA.
⁸ Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706 Wisconsin, USA.

Abstract

Homologs of the mutagenic Escherichia coli DNA polymerase V (pol V) are encoded by numerous pathogens and mobile elements. We have used Rum pol (RumA'2B), from the integrative conjugative element (ICE), R391, as a model mobile element-encoded polymerase (MEPol). The highly mutagenic Rum pol is transferred horizontally into a variety of recipient cells, including many pathogens. Moving between species, it is unclear if Rum pol can function on its own or requires activation by host factors. Here, we show that Rum pol biochemical activity requires the formation of a physical mutasomal complex, Rum Mut, containing RumA'2B-RecA-ATP, with RecA being donated by each recipient bacteria. For R391, Rum Mut specific activities in vitro and mutagenesis rates in vivo depend on the phylogenetic distance of host-cell RecA from E. coli RecA. Rum pol is a highly conserved and effective mobile catalyst of rapid evolution, with the potential to generate a broad mutational landscape that could serve to ensure bacterial adaptation in antibiotic-rich environments leading to the establishment of antibiotic resistance.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

DNA-Directed DNA Polymerase / metabolism
Escherichia coli* / metabolism
Mutagens*
Phylogeny
Rec A Recombinases* / metabolism

Substances

DNA-Directed DNA Polymerase
Mutagens
Rec A Recombinases
recA protein, E coli

Abstract

Publication types

MeSH terms

Substances

Grants and funding