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Localization of proteins bound to a replication origin of human DNA along the cell cycle
Author(s) -
Abdurashidova Gulnara,
Danailov Miltcho B.,
Ochem Alexander,
Triolo Gianluca,
Djeliova Vera,
Radulescu Sorina,
Vindigni Alessandro,
Riva Silvano,
Falaschi Arturo
Publication year - 2003
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/cdg404
Subject(s) - biology , dna replication , dna , genetics , cell cycle , origin recognition complex , microbiology and biotechnology , control of chromosome duplication , computational biology , replication (statistics) , cell cycle protein , dna binding protein , eukaryotic dna replication , cell , gene , transcription factor , virology
The proteins bound in vivo at the human lamin B2 DNA replication origin and their precise sites of binding were investigated along the cell cycle utilizing two novel procedures based on immunoprecipitation following UV irradiation with a pulsed laser light source. In G 1 , the pre‐replicative complex contains CDC6, MCM3, ORC1 and ORC2 proteins; of these, the post‐replicative complex in S phase contains only ORC2; in M phase none of them are bound. The precise nucleotide of binding was identified for the two ORC and the CDC6 proteins near the start sites for leading‐strand synthesis; the transition from the pre‐ to the post‐replicative complex is accompanied by a 17 bp displacement of the ORC2 protein towards the start site.