The DinI protein stabilizes RecA protein filaments.
When DinI is present at concentrations that are stoichiometric with those of RecA or somewhat greater, DinI has a substantial stabilizing effect on RecA filaments bound to DNA. Exchange of RecA between free and bound forms was almost entirely suppressed, and highly stable filaments were documented with several different experimental methods. DinI-mediated stabilization did not affect RecA-mediated ATP hydrolysis and LexA co-protease activities. Initiation of DNA strand exchange was affected in a DNA structure-dependent manner, whereas ongoing strand exchange was not affected. Destabilization of RecA filaments occurred as reported in earlier work but only when DinI protein was present at very high concentrations, generally superstoichiometric, relative to the RecA protein concentration. DinI did not facilitate RecA filament formation but stabilized the filaments only after they were formed. The interaction between the RecA protein and DinI was modulated by the C terminus of RecA. We discuss these results in the context of a new hypothesis for the role of DinI in the regulation of recombination and the SOS response.
Lusetti
Shelley L
SL
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706-1544, USA.
Voloshin
Oleg N
ON
Inman
Ross B
RB
Camerini-Otero
R Daniel
RD
Cox
Michael M
MM
D000255:Adenosine Triphosphate / Q000737:chemistry; D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism*; D005092:Exonucleases / Q000378:metabolism; D017353:Gene Deletion; D006868:Hydrolysis; D007477:Ions; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011938:Rec A Recombinases / Q000737:chemistry / Q000378:metabolism*; D011995:Recombination, Genetic; D020349:Surface Plasmon Resonance; D013997:Time Factors
D004277:DNA, Single-Stranded; C422844:DinI protein, E coli; D029968:Escherichia coli Proteins; D007477:Ions; D009841:Oligonucleotides; D000255:Adenosine Triphosphate; D004247:DNA; D011938:Rec A Recombinases; D005092:Exonucleases; D008274:Magnesium