Molecular characterization of Gluconobacter oxydans recA gene and its inhibitory effect on the function of the host wild-type recA gene.
A DNA fragment containing the recA gene of Gluconobacter oxydans was isolated and further characterized for its nucleotide sequence and ability to functionally complement various recA mutations. When expressed in an Escherichia coli recA host, the G. oxydans recA protein could efficiently function in homologous recombination and DNA damage repair. The recA gene's nucleotide sequence analysis revealed a protein of 344 amino acids with a molecular mass of 38 kDa. We observed an E. coli-like LexA repressor-binding site in the G. oxydans recA gene promoter region, suggesting that a LexA-like mediated response system may exist in G. oxydans. The expression of G. oxydans recA in E. coli RR1, a recA+ strain, surprisingly caused a remarkable reduction of the host wild-type recA gene function, whereas the expression of both Serratia marcescens recA and Pseudomonas aeruginosa recA gene caused only a slight inhibitory effect on function of the host wild-type recA gene product. Compared with the E. coli RecA protein, the identity of the amino acid sequence of G. oxydans RecA protein is much lower than those RecA proteins of both S. marcescens and Pseudomonas aeruginosa. This result suggests that the expression of another wild-type RecA could interfere with host wild-type recA gene's function, and the extent of such an interference is possibly correlated to the identity of the amino acid sequence between the two classes of RecA protein.
Liu
Y T
YT
Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, Republic of China. ytliu@ndmc1.ndmctsgh.edu.tw
Chao
D C
DC
Lee
F
F
Chen
C G
CG
Ji
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DD
D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016946:Gram-Negative Aerobic Rods and Cocci / Q000235:genetics*; D008969:Molecular Sequence Data; D011938:Rec A Recombinases / Q000037:antagonists & inhibitors / Q000235:genetics*; D011995:Recombination, Genetic; D013014:SOS Response, Genetics; D017422:Sequence Analysis, DNA; D014466:Ultraviolet Rays / Q000009:adverse effects
D011938:Rec A Recombinases