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Nucleic Acids Research, 1990, Vol. 18, No. 24 7425-7432
© 1990

Articles

Excision repair of UV-damaged plasmid DNA in Xenopus oocytes is mediated by DNA polymerase {alpha} (and/or {delta})

Jitendra K. Saxena, John B. Hays+ and Eric J. Ackerman*

Genetics and Biochemistry Branch, NIDDK, NIH Bethesda, MD 20892, USA

*To whom correspondence should be addressed

Received May 14, 1990. Revised November 13, 1990. Accepted November 13, 1990.

We studied DNA repair by injecting plasmids containing random pyrimidine dimers into Xenopus oocytes. We demonstrated excision repair by recovering plasmids and analyzing them with T4 UV endonuclease treatment and alkaline agarose gel electrophoresis. The mechanism for excision repair of these plasmids appears to be processive, rather than distributive, since repair occurs in an ‘all or none’ fashion. At less than 4–5 dimers/plasmid, nearly all repair occurs within 4–6 hours ({small tilde}1010 dimers repaired per oocyte); the oocyte, therefore, has abundant repair activity. Specific antibodies and inhibitors were used to determine enzymes involved in repair. We conclude that DNA polymerase {alpha} (and/or {delta}) is required because repair is inhibited by antibodies to human DNA polymerase {alpha}, as well as by aphidicolin, an inhibitor of polymerases {alpha} (and/or {delta}). Repair was not inhibited by hydroxyurea, cytosine ß-D-arabinofuranoside, or inhibitors of topoisomerase II (novobiocin). Oocyte repair does not activate semi-conservative DNA replication, nor is protein synthesis required. Photoreactivation cannot account for repair because dimer removal is independent of exogenous light.

+Present address: Department of Agricultural Chemistry, Oregon State University, Corvallis, OR 97331, USA


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N. Oda, J. K. Saxena, T. M. Jenkins, R. Prasad, S. H. Wilson, and E. J. Ackerman
DNA Polymerases alpha and beta Are Required for DNA Repair in an Efficient Nuclear Extract from Xenopus Oocytes
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T. Jenkins, J. Saxena, A Kumar, S. Wilson, and E. Ackerman
DNA polymerase beta and DNA synthesis in Xenopus oocytes and in a nuclear extract
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