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
|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.
|Permanent link:||http://hdl.handle.net/10197/4998||Date:||10-Aug-2012||Online since:||2013-11-28T14:55:49Z||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||Journal:||Journal of Biological Chemistry||Volume:||287||Issue:||40||Start page:||33327||End page:||33338||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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