Characterization of RecA424 and RecA670 proteins from Deinococcus radiodurans.
RecA protein is considered to be the most important participant in the radiation resistance of Deinococcus radiodurans. However, it is still unclear how RecA contributes to the resistance. In this study, we identified a new recA mutation (recA424) in the DNA-repair deficient mutant strain KI696, the phenotype of which is remarkably different from mutant strain rec30 carrying recA670. The properties of the gene products from the recA mutants were compared. recA424 could not complement the deficiency in Escherichia coli RecA, as found for recA670. In vitro, neither RecA424 nor RecA670 could promote DNA strand exchange under conditions in which wild-type RecA promoted the reaction, indicating that both RecA424 and Rec670 are defective in recombination activity. RecA424 promoted the autocleavage reaction of LexA in vitro, whereas RecA670 did not. The intracellular LexA level in KI696 was decreased following gamma-irradiation. However, the LexA level in strain rec30 was constant irrespective of irradiation. These results indicate that RecA424 retains co-protease activity, whereas RecA670 does not. While strain rec30 is extremely radiation sensitive, strain KI696 is only slightly sensitive. Together, these observations suggest that the co-protease activity rather than the recombination activity of RecA contributes to radiation resistance in D. radiodurans.
Satoh
Katsuya
K
Biotechnology Laboratory, Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute, Takasaki 370-1292, Japan.
Narumi
Issay
I
Kikuchi
Masahiro
M
Kitayama
Shigeru
S
Yanagisawa
Tadashi
T
Yamamoto
Kazuo
K
Watanabe
Hiroshi
H
D001419:Bacteria / Q000201:enzymology; D001426:Bacterial Proteins / Q000378:metabolism*; D004260:DNA Repair / Q000502:physiology*; D005720:Gamma Rays; D005816:Genetic Complementation Test; D009154:Mutation; D010447:Peptide Hydrolases / Q000378:metabolism; D011836:Radiation Tolerance; D011938:Rec A Recombinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011995:Recombination, Genetic; D012697:Serine Endopeptidases / Q000378:metabolism*; D014170:Transformation, Genetic; D000099160:LexA Repressor Protein
D001426:Bacterial Proteins; D010447:Peptide Hydrolases; D011938:Rec A Recombinases; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein