Characterization of a UV Endonuclease Gene from the Fisson Yeast Schizosaccharomyces Pombe and Its Bacterial Homolog

Abstract

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.