Header

UZH-Logo

Maintenance Infos

Cell cycle-dependent dynamic association of cyclin/Cdk complexes with human DNA replication proteins.


Frouin, I; Montecucco, A; Biamonti, G; Hübscher, U; Spadari, S; Maga, G (2002). Cell cycle-dependent dynamic association of cyclin/Cdk complexes with human DNA replication proteins. The EMBO Journal, 21(10):2485-2495.

Abstract

We have previously described the isolation of a replication competent (RC) complex from calf thymus, containing DNA polymerase alpha, DNA polymerase delta and replication factor C. Here, we describe the isolation of the RC complex from nuclear extracts of synchronized HeLa cells, which contains DNA replication proteins associated with cell-cycle regulation factors like cyclin A, cyclin B1, Cdk2 and Cdk1. In addition, it contains a kinase activity and DNA polymerase activities able to switch from a distributive to a processive mode of DNA synthesis, which is dependent on proliferating cell nuclear antigen. In vivo cross-linking of proteins to DNA in synchronized HeLa cells demonstrates the association of this complex to chromatin. We show a dynamic association of cyclins/Cdks with the RC complex during the cell cycle. Indeed, cyclin A and Cdk2 associated with the complex in S phase, and cyclin B1 and Cdk1 were present exclusively in G(2)/M phase, suggesting that the activity, as well the localization, of the RC complex might be regulated by specific cyclin/Cdk complexes.

Abstract

We have previously described the isolation of a replication competent (RC) complex from calf thymus, containing DNA polymerase alpha, DNA polymerase delta and replication factor C. Here, we describe the isolation of the RC complex from nuclear extracts of synchronized HeLa cells, which contains DNA replication proteins associated with cell-cycle regulation factors like cyclin A, cyclin B1, Cdk2 and Cdk1. In addition, it contains a kinase activity and DNA polymerase activities able to switch from a distributive to a processive mode of DNA synthesis, which is dependent on proliferating cell nuclear antigen. In vivo cross-linking of proteins to DNA in synchronized HeLa cells demonstrates the association of this complex to chromatin. We show a dynamic association of cyclins/Cdks with the RC complex during the cell cycle. Indeed, cyclin A and Cdk2 associated with the complex in S phase, and cyclin B1 and Cdk1 were present exclusively in G(2)/M phase, suggesting that the activity, as well the localization, of the RC complex might be regulated by specific cyclin/Cdk complexes.

Statistics

Citations

39 citations in Web of Science®
39 citations in Scopus®
Google Scholar™

Altmetrics

Downloads

138 downloads since deposited on 11 Feb 2008
13 downloads since 12 months
Detailed statistics

Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:05 Vetsuisse Faculty > Department of Molecular Mechanisms of Disease
07 Faculty of Science > Department of Molecular Mechanisms of Disease
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:15 May 2002
Deposited On:11 Feb 2008 12:18
Last Modified:06 Dec 2017 13:00
Publisher:European Molecular Biology Organization ; Nature Publishing Group
ISSN:0261-4189
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1093/emboj/21.10.2485
PubMed ID:12006500

Download

Download PDF  'Cell cycle-dependent dynamic association of cyclin/Cdk complexes with human DNA replication proteins.'.
Preview
Filetype: PDF
Size: 332kB
View at publisher