Characterization of a UV endonuclease gene from the fission yeast Schizosaccharomyces pombe and its bacterial homolog.
From the fission yeast Schizosaccharomyces pombe, a cDNA fragment was isolated, which confers UV resistance on repair deficient Escherichia coli host cells. The cloned cDNA encodes a protein of 68,815 Da, which has a 36.6% identity of amino acid sequence with the previously identified 74 kDa UV endonuclease of the filamentous fungus Neurospora crassa. Analysis of several truncated gene constructs shows that only the C-terminal two thirds region, which has 54% identity of amino acid sequence with the C-terminal region of the Neurospora homolog, is necessary for complementing activity of UV-sensitivity in the E. coli host cells. Purified recombinant protein from E. coli host cells incises both UV-induced cyclobutane pyrimidine dimers and (6-4) photoproducts at the sites immediately 5' to the DNA damage in the same fashion as the Neurospora protein. Furthermore, a bacterial homologous sequence was isolated from Bacillus subtilis and shows a similar complementing activity of UV sensitivity in E. coli host cells, indicating a wide distribution of this alternative excision repair mechanism in life.
Takao
M
M
Department of Neurochemistry and Molecular Biology, Institute of Development, Ageing and Cancer, Tohoku University, Sendai, Japan.
Yonemasu
R
R
Yamamoto
K
K
Yasui
A
A
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular; D004260:DNA Repair; D018076:DNA, Complementary / Q000235:genetics; D004706:Endodeoxyribonucleases / Q000235:genetics*; D005798:Genes, Bacterial; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000235:genetics; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D029702:Schizosaccharomyces pombe Proteins; D004706:Endodeoxyribonucleases; C105073:uve1 protein, S pombe