Transposition of insertion sequence IS256Bsu1 in Bacillus subtilis 168 is strictly dependent on recA.
We developed an insertion sequence transposition detection system called the "jumping cat assay" and applied it to the Bacillus subtilis chromosome using IS256Bsu1 derived from B. subtilis natto. The high frequency of transposition enabled us to explore host factors; combining the assay and genetic analyses revealed that recA is essential for the transposition of IS256Bsu1. Detailed analyses using various domain mutants of recA demonstrated that this essentiality is not related to the function of recA in homologous recombination. Instead, the ATP binding and hydrolysis function seemed to be crucial for IS transposition. To elucidate the role of recA, we focused on the muB gene of the enterobacteriophage Mu. Based on information from the NCBI Conserved Domain Database, both MuB and RecA belong to the P-loop dNTPase superfamily. Further experiments revealed that muB complements the transposition-defective phenotype of a recA deletant, although it could not rescue UV sensitivity. These results suggest that recA shares a common function with muB that helps the transposition of IS256Bsu1 in B. subtilis.
Department of Bioscience, Tokyo University of Agriculture.
Motohiro
M
Akashi
Department of Bioscience, Tokyo University of Agriculture.
Shota
S
Harada
Department of Bioscience, Tokyo University of Agriculture.
Syunsuke
S
Moki
Department of Bioscience, Tokyo University of Agriculture.
Yuki
Y
Okouji
Department of Bioscience, Tokyo University of Agriculture.
Kiwamu
K
Takahashi
Central Research Institute, Mitsukan Group Co., Ltd.
Shigeki
S
Kada
Central Research Institute, Mitsukan Group Co., Ltd.
Keigo
K
Yamagami
Department of Life Science, College of Science, Rikkyo (St Paul's) University.
Yasuhiko
Y
Sekine
Department of Bioscience, Tokyo University of Agriculture.
Satoru
S
Watanabe
Department of Bioscience, Tokyo University of Agriculture.
Taku
T
Chibazakura
Department of Bioscience, Tokyo University of Agriculture.
Hirofumi
H
Yoshikawa
D000255:Adenosine Triphosphate / Q000378:metabolism; D001412:Bacillus subtilis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000235:genetics; D059765:Homologous Recombination; D009154:Mutation; D011485:Protein Binding; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C069215:MuB protein, Enterobacteria phage Mu; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases
ATPase domain; Bacillus subtilis; insertion sequence; muB; recA