DNA Polymerases α and β Are Required for DNA Repair in an Efficient Nuclear Extract from Xenopus Oocytes

Xenopus oocytes and an oocyte nuclear extract efficiently repair the bulky DNA lesions cyclobutane pyrimidine dimers, (6-4) photoproducts, and N-acetoxy-2-aminofluorene (AAF) adducts by an excision repair mechanism. Nearly all (>95%) of the input damaged DNA was repaired within 5 h in both inject...

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Bibliographic Details
Published in:The Journal of biological chemistry 1996-06, Vol.271 (23), p.13816-13820
Main Authors: Oda, Naoko, Saxena, Jitendra K., Jenkins, Timothy M., Prasad, Rajendra, Wilson, Samuel H., Ackerman, Eric J.
Format: Article
Language:English
Subjects:
Animals
Antibodies - pharmacology
Aphidicolin - pharmacology
Cell Nucleus - metabolism
DNA Polymerase I - antagonists & inhibitors
DNA Polymerase I - metabolism
DNA Polymerase II - antagonists & inhibitors
DNA Polymerase II - metabolism
DNA Repair - physiology
Enzyme Inhibitors - pharmacology
Female
Freshwater
Humans
In Vitro Techniques
Oocytes - metabolism
Pyrimidine Dimers - metabolism
Pyrimidine Dimers - radiation effects
Ultraviolet Rays
Xenopus
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Summary:Xenopus oocytes and an oocyte nuclear extract efficiently repair the bulky DNA lesions cyclobutane pyrimidine dimers, (6-4) photoproducts, and N-acetoxy-2-aminofluorene (AAF) adducts by an excision repair mechanism. Nearly all (>95%) of the input damaged DNA was repaired within 5 h in both injected cells and extracts with no significant incorporation of label into control undamaged DNA. Remarkably, more than 1010 cyclobutane pyrimidine dimers or (6-4) photoproducts are repaired/nuclei. The extracts are free from nuclease activity, and repair is independent of exogenous light. Both the high efficiency and DNA polymerase requirements of this system appear to be different from extracts derived from human cells. We demonstrated a requirement for DNA polymerases α and β in repair of both photoproducts and AAF by inhibiting repair with several independent antibodies specific to either DNA polymerases α or β and then restoring repair by adding the appropriate purified polymerase. Repair is inhibited by aphidicolin at concentrations specific for blocking DNA polymerase α and dideoxynucleotide triphosphates at concentrations specific for inhibiting DNA polymerase β.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.271.23.13816