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Biochemistry. 1998 Aug 18;37(33):11599-604.

Expression, purification, and characterization of ultraviolet DNA endonuclease from Schizosaccharomyces pombe.

Author information

1
Department of Biochemistry, Division of Cancer Biology, Radiation and Oncology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.

Abstract

Ultraviolet damage endonuclease (UVDE) is a 68.7 kDa DNA repair enzyme of Schizosaccharomyces pombe that recognizes cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4 PPs). UVDE is thought to initiate the first step in an alternative excision repair pathway for removal of UV light-induced DNA damage. We have overexpressed Delta228-UVDE, an active truncated form of UVDE, and have purified this protein to apparent homogeneity. We have characterized purified Delta228-UVDE with respect to its physical properties, divalent cation requirements, and kinetic parameters on oligodeoxynucleotide substrates containing a single CPD. DNA strand cleavage analysis indicates that both full-length UVDE and Delta228-UVDE incise the CPD-containing strand immediately 5' to the lesion. These results provide further insight into the UVDE-mediated alternative excision repair pathway.

PMID:
9708997
DOI:
10.1021/bi981008c
[Indexed for MEDLINE]

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