Amplified UvrA protein can ameliorate the ultraviolet sensitivity of an Escherichia coli recA mutant.
When a recA strain of Escherichia coli was transformed with the multicopy plasmid pSF11 carrying the uvrA gene of E. coli, its extreme ultraviolet (UV) sensitivity was decreased. The sensitivity of the lexA1 (Ind(-)) strain to UV was also decreased by pSF11. The recA cells expressing Neurospora crassa UV damage endonuclease (UVDE), encoding UV-endonuclease, show UV resistance. On the other hand, only partial amelioration of UV sensitivity of the recA strain was observed in the presence of the plasmid pNP10 carrying the uvrB gene. Host cell reactivation of UV-irradiated lambda phage in recA cells with pSF11 was as efficient as that in wild-type cells. Using an antibody to detect cyclobutane pyrimidine dimers, we found that UV-irradiated recA cells removed dimers from their DNA more rapidly if they carried pSF11 than if they carried a vacant control plasmid. Using anti-UvrA antibody, we observed that the expression level of UvrA protein was about 20-fold higher in the recA strain with pSF11 than in the recA strain without pSF11. Our results were consistent with the idea that constitutive level of UvrA protein in the recA cells results in constitutive levels of active UvrABC nuclease which is not enough to operate full nucleotide excision repair (NER), thus leading to extreme UV sensitivity.
Kiyosawa
K
K
Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai 980-8578, Japan.
Tanaka
M
M
Matsunaga
T
T
Nikaido
O
O
Yamamoto
K
K
D000251:Adenosine Triphosphatases / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D010582:Bacteriophage lambda / Q000254:growth & development / Q000528:radiation effects; D004249:DNA Damage; D004265:DNA Helicases / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D004260:DNA Repair; D004279:DNA, Viral / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D004720:Endonucleases / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000528:radiation effects* / Q000821:virology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics; D011740:Pyrimidine Dimers / Q000378:metabolism; D011938:Rec A Recombinases / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000502:physiology; D013014:SOS Response, Genetics / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics / Q000502:physiology; D014466:Ultraviolet Rays; D014775:Virus Activation; D000099160:LexA Repressor Protein
D000251:Adenosine Triphosphatases; D001426:Bacterial Proteins; D004265:DNA Helicases; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D004720:Endonucleases; D029968:Escherichia coli Proteins; D005656:Fungal Proteins; D011740:Pyrimidine Dimers; D011938:Rec A Recombinases; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D000099160:LexA Repressor Protein; C058093:UvrB protein, E coli; C445669:UVDE protein, Neurospora crassa; C068631:UvrA protein, E coli