PrfA protein of Bacillus species: prediction and demonstration of endonuclease activity on DNA.
The prfA gene product of Gram-positive bacteria is unusual in being implicated in several cellular processes; cell wall synthesis, chromosome segregation, and DNA recombination and repair. However, no homology of PrfA with other proteins has been evident. Here we report a structural relationship between PrfA and the restriction enzyme PvuII, and thereby produce models that predict that PrfA binds DNA. Indeed, wild-type Bacillus stearothermophilus PrfA, but not a catalytic site mutant, nicked one strand of supercoiled plasmid templates leaving 5'-phosphate and 3'-hydroxyl termini. This activity, much lower on linear or relaxed circular double-stranded DNA or on single-stranded DNA, is consistent with a role for this protein in chromosome segregation, DNA recombination, or DNA repair.
Rigden
Daniel J
DJ
National Centre of Genetic Resources and Biotechnology, Cenargen/Embrapa, Brasília, Brazil, D.F. 70770-900. daniel@cenargen.embrapa.br
Setlow
Peter
P
Setlow
Barbara
B
Bagyan
Irina
I
Stein
Richard A
RA
Jedrzejas
Mark J
MJ
D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics / Q000378:metabolism*; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics; D029968:Escherichia coli Proteins; D008565:Membrane Proteins / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010454:Peptide Termination Factors; D010802:Phylogeny; D011487:Protein Conformation; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C096452:FtsK protein, E coli; D008565:Membrane Proteins; D010454:Peptide Termination Factors; D015534:Trans-Activators; D004247:DNA; D004262:DNA Restriction Enzymes; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific