Aoufouchi, Said and Patrick, Tina and Lindsay, Howard D. and Shall, Sydney and Ford, Christopher C. (1997) Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts. FEBS Journal, 247 (2). pp. 518-525. ISSN 1742-464X
Full text not available from this repository.Abstract
We have analysed the recircularisation of plasmid DNA, cut with two different endonucleases to generate non-homologous DNA ends, in extracts of unfertilised eggs and oocytes of Xenopus. We found that the capacity to join non-homologous DNA ends, generating diagnostic covalently closed monomer circles, appeared during oocyte maturation at the time of germinal vesicle breakdown. This enzyme function was post-translationally activated in oocyte extracts incubated with unfertilised egg extract containing active cdc2kyclin B, or by incubation with purified cdc2/cyclin B. Dephosphorylation of egg proteins by alkaline phosphatase inhibited the ability to join non-homologous DNA endr. We show that most linear non-homologous DNA ends repaired to form closed-circular supercoiled monomers, are joined without loss of nucleotides. Following partial purification, the activity was inhibited by inhibitors of poly(ADP-Rib) polymerase, an enzyme that is inactive in oocytes, but phosphorylated and activated during maturation. Competitive inhibition of poly(ADP-Rib) polymerase by > 50 pM 3-aminobenzamide prevented the joining of both matched and non-homologous DNA ends. We conclude that post-translational phosphorylation provides one route by which end-joining of non-homologous DNA can be regulated.
| Item Type: | Article |
|---|---|
| Journal or Publication Title: | FEBS Journal |
| Uncontrolled Keywords: | non-homologous recombination ; Xenopus extracts ; poly(ADP-ribose) polymerase ; cyclindependent kinase ; post-translational control. |
| Subjects: | Q Science > QH Natural history > QH301 Biology |
| Departments: | Faculty of Health and Medicine > Medicine |
| ID Code: | 28223 |
| Deposited By: | Dr Howard Lindsay |
| Deposited On: | 17 Nov 2009 13:07 |
| Refereed?: | Yes |
| Published?: | Published |
| Last Modified: | 26 Jul 2012 16:46 |
| Identification Number: | |
| URI: | http://eprints.lancs.ac.uk/id/eprint/28223 |
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