Replacement of the lysine residue in the consensus ATP-binding sequence of the AddA subunit of AddAB drastically affects chromosomal recombination in transformation and transduction of Bacillus subtilis.
The ATP-dependent deoxyribonuclease enzyme complex (AddAB) of Bacillus subtilis possesses two consensus ATP-binding sequences, located in the N-terminal region of both subunits. The highly conserved lysine residues in both consensus ATP-binding sequences were replaced by glycine, resulting in the mutant enzyme complexes AddAB-A-K36G (AddA*B) and AddAB-B-K14G (AddAB*). The mutation in subunit AddA reduced DNA repair and chromosomal transformation, and abolished bacteriophage PBS1-mediated transduction. This mutation also resulted in a complete loss of the ATP-dependent exonuclease and helicase activity. In contrast, the mutation in subunit AddB had only marginal effects. The recF and addAB genes are not required for transformation with plasmid DNA, but have overlapping activities in transformation with chromosomal DNA. By contrast to RecF, the AddAB enzyme is essential for PBS1-mediated transduction. However, recF has a more important function with respect to DNA repair than addAB.
Haijema
B J
BJ
Department of Genetics, University of Groningen, Haren, The Netherlands.
Noback
M
M
Hesseling
A
A
Kooistra
J
J
Venema
G
G
Meima
R
R
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D001665:Binding Sites / Q000235:genetics; D016384:Consensus Sequence; D004265:DNA Helicases; D004260:DNA Repair; D005090:Exodeoxyribonucleases; D005092:Exonucleases; D008239:Lysine / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D000255:Adenosine Triphosphate; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C080948:AddA protein, Bacillus subtilis; C080949:AddB protein, Bacillus subtilis; D004265:DNA Helicases; D008239:Lysine