TAILIEUCHUNG - Báo cáo khoa học: Distinctive activities of DNA polymerases during human DNA replication

The contributions of human DNA polymerases (pols)a, dande during S-phase progression were studied in order to elaborate how these enzymes co-ordinate their functions during nuclear DNA replication. Pol d was three to four times more intensely UV cross-linked to nascent DNA in late compared with early S phase, whereas the cross-linking of pols aande remained nearly constant throughout the S phase. | iFEBS Journal Distinctive activities of DNA polymerases during human DNA replication Anna K. Rytkonen1 2 Markku Vaara2 Tamar Nethanel3 Gabriel Kaufmann3 Raija Sormunen4 Esa Laara5 Heinz-Peter Nasheuer6 Amal Rahmeh7 Marietta Y. W. T. Lee7 Juhani E. Syvaoja2 and Helmut Pospiech1 1 Biocenter Oulu and Department of Biochemistry University of Oulu Finland 2 Department of Biology University of Joensuu Finland 3 Department of Biochemistry TelAviv University Israel 4 Biocenter Oulu and Department of Pathology University of Oulu Finland 5 Department of MathematicalSciences University of Oulu Finland 6 NationalUniversity of Ireland Department of Biochemistry CellCycle ControlLaboratory Galway Ireland 7 Department of Biochemistry and Molecular Biology New York MedicalCollege Valhalla NY USA Keywords cell cycle DNA polymerase DNA replication electron microscopy UV crosslinking Correspondence H. Pospiech Department of Biochemistry PO Box 3000 FIN-90014 University of Oulu Finland Fax 358 8 553 1141 Tel 358 8 553 1155 E-mail Received 20 March 2006 revised 3 May 2006 accepted 5 May 2006 doi The contributions of human DNA polymerases pols a Ỗ and e during S-phase progression were studied in order to elaborate how these enzymes co-ordinate their functions during nuclear DNA replication. Pol Ỗ was three to four times more intensely UV cross-linked to nascent DNA in late compared with early S phase whereas the cross-linking of pols a and e remained nearly constant throughout the S phase. Consistently the chromatin-bound fraction of pol Ỗ unlike pols a and e increased in the late S phase. Moreover pol Ỗ neutralizing antibodies inhibited replicative DNA synthesis most efficiently in late S-phase nuclei whereas antibodies against pol e were most potent in early S phase. Ultrastructural localization of the pols by immuno-electron microscopy revealed pol e to localize predominantly to ring-shaped clusters at electron-dense regions of

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