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T‐antigen interactions with chromatin and p53 during the cell cycle in extracts from Xenopus eggs
Author(s) -
Vassetzky Yegor S.,
Tchang Françoise,
Fanning Ellen,
Méchali Marcel
Publication year - 1999
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19991101)75:2<288::aid-jcb11>3.0.co;2-7
Subject(s) - chromatin , microbiology and biotechnology , biology , dna replication , cell cycle , xenopus , proliferating cell nuclear antigen , eukaryotic dna replication , nuclear matrix , antigen , control of chromosome duplication , origin recognition complex , nuclear transport , nuclear membrane , dna , cell , cell nucleus , genetics , nucleus , gene
The role of SV40 large tumor T‐antigen in replication of viral DNA is well established, but it is still unclear how T‐antigen triggers cellular replication and cell transformation in non‐permissive cells. Here, we used Xenopus egg extracts which reproduce most nuclear events linked to the cell cycle in vitro to analyze its interaction with genomic chromatin during the cell cycle. We show that T‐antigen associates with chromatin before the nuclear membrane formation, and further demonstrate that the nuclear membrane is not necessary for its import into the nucleus. We show that the interaction of T‐antigen with the endogenous chromatin does not occur at replication foci nor at RPA pre‐replication centers. Immunoprecipitations as well as sucrose gradient experiments, indicate that the endogenous pool of p53 interacts with T‐antigen. In addition, a transient association of both proteins with the nuclear matrix is observed during the ongoing DNA synthesis. These data are discussed in view of the T‐antigen and p53 activity during the cell cycle. J. Cell. Biochem. 75:288–299, 1999. © 1999 Wiley‐Liss, Inc.