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Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts.

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Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts. / Aoufouchi, Said; Patrick, Tina; Lindsay, Howard D.; Shall, Sydney; Ford, Christopher C.

In: FEBS Journal, Vol. 247, No. 2, 07.1997, p. 518-525.

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Aoufouchi, Said ; Patrick, Tina ; Lindsay, Howard D. ; Shall, Sydney ; Ford, Christopher C. / Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts. In: FEBS Journal. 1997 ; Vol. 247, No. 2. pp. 518-525.

Bibtex

@article{0e78bea328fc4b11b8fa01efc7938235,
title = "Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts.",
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.",
keywords = "non-homologous recombination, Xenopus extracts, poly(ADP-ribose) polymerase, cyclindependent kinase, post-translational control.",
author = "Said Aoufouchi and Tina Patrick and Lindsay, {Howard D.} and Sydney Shall and Ford, {Christopher C.}",
year = "1997",
month = jul,
doi = "10.1111/j.1432-1033.1997.00518.x",
language = "English",
volume = "247",
pages = "518--525",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell Publishing Ltd",
number = "2",

}

RIS

TY - JOUR

T1 - Post-translational activation of non-homologous DNA end-joining in Xenopus oocyte extracts.

AU - Aoufouchi, Said

AU - Patrick, Tina

AU - Lindsay, Howard D.

AU - Shall, Sydney

AU - Ford, Christopher C.

PY - 1997/7

Y1 - 1997/7

N2 - 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.

AB - 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.

KW - non-homologous recombination

KW - Xenopus extracts

KW - poly(ADP-ribose) polymerase

KW - cyclindependent kinase

KW - post-translational control.

U2 - 10.1111/j.1432-1033.1997.00518.x

DO - 10.1111/j.1432-1033.1997.00518.x

M3 - Journal article

VL - 247

SP - 518

EP - 525

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 2

ER -