title: Dimer exchange and cleavage specificity of the DNA damage response protein UmuD.
abstract: The cellular response to DNA damage in Escherichia coli is controlled in part by the activity of the umuD gene products. The full-length dimeric UmuD(2) is the initial product that is expressed shortly after the induction of the SOS response and inhibits bacterial mutagenesis, allowing for error-free repair to occur. Over time, the slow auto-cleavage of UmuD(2) to UmuD'(2) promotes mutagenesis to ensure cell survival. The intracellular levels of UmuD(2) and UmuD'(2) are further regulated by degradation in vivo, returning the cell to a non-mutagenic state. To further understand the dynamic regulatory roles of the umuD gene products, we monitored the kinetics of exchange and cleavage of the UmuD(2) and UmuD'(2) homodimers as well as of the UmuDD' heterodimer under equilibrium conditions. We found that the heterodimer is the preferred but not exclusive protein form, and that both the heterodimer and homodimers exhibit slow exchange kinetics which is further inhibited in the presence of interacting partner DinB. In addition, the heterodimer efficiently cleaves to form UmuD'(2). Together, this work reveals an intricate UmuD lifecycle that involves dimer exchange and cleavage in the regulation of the DNA damage response.
authors.affiliation: Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, 02115, USA.
authors.forename: Jaylene N
authors.initials: JN
authors.lastname: Ollivierre
authors.forename: Jacquelyn L
authors.initials: JL
authors.lastname: Sikora
authors.forename: Penny J
authors.initials: PJ
authors.lastname: Beuning
mesh_terms: D004249:DNA Damage; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D016296:Mutagenesis; D055503:Protein Multimerization / Q000502:physiology*
chemical_list: D004269:DNA, Bacterial; C094399:DinB protein, E coli; D029968:Escherichia coli Proteins; D004259:DNA-Directed DNA Polymerase; C055218:UmuD protein, E coli