Segregation of Replicative DNA Polymerases during S Phase: DNA POLYMERASE ε, BUT NOT DNA POLYMERASES α/δ, ARE ASSOCIATED WITH LAMINS THROUGHOUT S PHASE IN HUMAN CELLS

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Title: Segregation of Replicative DNA Polymerases during S Phase: DNA POLYMERASE ε, BUT NOT DNA POLYMERASES α/δ, ARE ASSOCIATED WITH LAMINS THROUGHOUT S PHASE IN HUMAN CELLS
Authors: Vaara, M.
Itkonen, H.
Hillukkala, T.
et al.
Permanent link: http://hdl.handle.net/10197/4998
Date: 10-Aug-2012
Abstract: DNA polymerases (Pol) α, δ, and ϵ replicate the bulk of chromosomal DNA in eukaryotic cells, Pol ϵ being the main leading strand and Pol δ the lagging strand DNA polymerase. By applying chromatin immunoprecipitation (ChIP) and quantitative PCR we found that at G1/S arrest, all three DNA polymerases were enriched with DNA containing the early firing lamin B2 origin of replication and, 2 h after release from the block, with DNA containing the origin at the upstream promoter region of the MCM4 gene. Pol α, δ, and ϵ were released from these origins upon firing. All three DNA polymerases, Mcm3 and Cdc45, but not Orc2, still formed complexes in late S phase. Reciprocal ChIP of the three DNA polymerases revealed that at G1/S arrest and early in S phase, Pol α, δ, and ϵ were associated with the same nucleoprotein complexes, whereas in late S phase Pol ϵ and Pol α/δ were largely associated with distinct complexes. At G1/S arrest, the replicative DNA polymerases were associated with lamins, but in late S phase only Pol ϵ, not Pol α/δ, remained associated with lamins. Consistently, Pol ϵ, but not Pol δ, was found in nuclear matrix fraction throughout the cell cycle. Therefore, Pol ϵ and Pol α/δ seem to pursue their functions at least in part independently in late S phase, either by physical uncoupling of lagging strand maturation from the fork progression, or by recruitment of Pol δ, but not Pol ϵ, to post-replicative processes such as translesion synthesis or post-replicative repair.
Type of material: Journal Article
Publisher: American Society for Biochemistry and Molecular Biology
Copyright (published version): 2012 American Society for Biochemistry and Molecular Biology
Keywords: Chromatin;Chromatin Immunoprecipitation (ChiP);DNA Polymerase;DNA Replication;DNA-Protein Interaction
DOI: 10.1074/jbc.M112.357996
Language: en
Status of Item: Peer reviewed
Appears in Collections:SBI Research Collection

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