About: Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities

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About: Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities Goto Sponge NotDistinct Permalink

An Entity of Type : http://linked.opendata.cz/ontology/domain/vavai/Vysledek, within Data Space : linked.opendata.cz associated with source document(s)

Attributes	Values
rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
rdfs:seeAlso	http://www.sciencedirect.com/science/article/pii/S1568786411000772
Description	The error-free repair of double-strand DNA breaks by homologous recombination (HR) ensures genomic stability using undamaged homologous sequence to copy genetic information. While some of the aspects of the initial steps of HR are understood, the molecular mechanisms underlying events downstream of the D-loop formation remain unclear. Therefore, we have reconstituted D-loop-based in vitro recombination-associated DNA repair synthesis assay and tested the efficacy of polymerases Pol δ and Pol η to extend invaded primer, and the ability of three helicases (Mph1, Srs2 and Sgs1) to displace this extended primer. Both Pol δ and Pol η extended up to 50% of the D-loop substrate, but differed in product length and dependency on proliferating cell nuclear antigen (PCNA). Mph1, but not Srs2 or Sgs1, displaced the extended primer very efficiently, supporting putative role of Mph1 in promoting the synthesis-dependent strand-annealing pathway. The experimental system described here can be employed to increase our understanding of HR events following D-loop formation, as well as the regulatory mechanisms involved. The error-free repair of double-strand DNA breaks by homologous recombination (HR) ensures genomic stability using undamaged homologous sequence to copy genetic information. While some of the aspects of the initial steps of HR are understood, the molecular mechanisms underlying events downstream of the D-loop formation remain unclear. Therefore, we have reconstituted D-loop-based in vitro recombination-associated DNA repair synthesis assay and tested the efficacy of polymerases Pol δ and Pol η to extend invaded primer, and the ability of three helicases (Mph1, Srs2 and Sgs1) to displace this extended primer. Both Pol δ and Pol η extended up to 50% of the D-loop substrate, but differed in product length and dependency on proliferating cell nuclear antigen (PCNA). Mph1, but not Srs2 or Sgs1, displaced the extended primer very efficiently, supporting putative role of Mph1 in promoting the synthesis-dependent strand-annealing pathway. The experimental system described here can be employed to increase our understanding of HR events following D-loop formation, as well as the regulatory mechanisms involved. (en)
Title	Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities (en)
skos:prefLabel	Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities Reconstitution of DNA repair synthesis in vitro and the role of polymerase and helicase activities (en)
skos:notation	RIV/00159816:_____/11:#0000764!RIV12-MZ0-00159816
http://linked.open...avai/riv/aktivita	O P
http://linked.open...avai/riv/aktivity	O, P(GA301/09/1917), P(GD203/09/H046), P(LC06030), P(ME10048)
http://linked.open...iv/cisloPeriodika	6
http://linked.open...vai/riv/dodaniDat	2012
http://linked.open...aciTvurceVysledku	Krejčí, Lumír
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Fakultní nemocnice u sv.Anny v Brně
http://linked.open...dnocenehoVysledku	225974
http://linked.open...ai/riv/idVysledku	RIV/00159816:_____/11:#0000764
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	DNA repair, Recombination, DNA synthesis, Replication, Mph1, Srs2 (en)
http://linked.open.../riv/klicoveSlovo	Recombination DNA repair DNA synthesis Mph1 Replication Srs2
http://linked.open...odStatuVydavatele	NL - Nizozemsko
http://linked.open...ontrolniKodProRIV	[E2EC99EDCE9B]
http://linked.open...i/riv/nazevZdroje	DNA Repair
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	1 (xsd:int)
http://linked.open...cetTvurcuVysledku	4 (xsd:int)
http://linked.open...vavai/riv/projekt	Biomolecular centre Biochemistry on the crossroad from in silico to in vitro Resolution of replication-recombination DNA intermediates and its role in genomic instability The role of post-translational modification in DNA repair and recombination: Meiosis, SUMOylation and Zip3 protein
http://linked.open...UplatneniVysledku	2011
http://linked.open...v/svazekPeriodika	10
http://linked.open...iv/tvurceVysledku	Burkovics, P. Haracska, L. Krejčí, Lumír Šebesta, M.
issn	1568-7864
number of pages	10 (xsd:int)
http://bibframe.org/vocab/doi	10.1016/j.dnarep.2011.03.003

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