Mechanism of action of Micrococcus luteus. gamma. -endonuclease
Micrococcus luteus extracts contain ..gamma..-endonuclease, a Mg/sup 2 +/-independent endonuclease that cleaves ..gamma..-irradiated DNA. This enzyme has been purified approximately 1000-fold, and the purified enzyme was used to study its substrate specificity and mechanism of action. ..gamma..-Endonuclease cleaves DNA containing either thymine glycols, urea residues, or apurinic sites but not undamaged DNA or DNA containing reduced apurinic sites. The enzyme has both N-glycosylase activity that releases thymine glycol residues from OsO/sub 4/-treated DNA and an associated apurinic endonuclease activity. The location and nature of the cleavage site produced has been determined with DNA sequencing techniques. ..gamma..-Endonuclease cleaves DNA containing thymine glycols or apurinic sites immediately 3' to the damaged or missing base. Cleavage results in a 5'-phosphate terminus and a 3' baseless sugar residue. Cleavage sites can be converted to primers for DNA polymerase I by subsequent treatment with Escherichia coli exonuclease III. The mechanism of action of ..gamma..-endonuclease and its substrate specificity are very similar to those identified for E. coli endonuclease III.
- Research Organization:
- Dana-Farber Cancer Institute, Boston, MA
- OSTI ID:
- 5397910
- Journal Information:
- Biochemistry; (United States), Journal Name: Biochemistry; (United States) Vol. 26:20; ISSN BICHA
- Country of Publication:
- United States
- Language:
- English
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