Localization of proteins bound to a replication origin of human DNA along the cell cycle.
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.
Abdurashidova
Gulnara
G
International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34012 Trieste, Italy.
Danailov
Miltcho B
MB
Ochem
Alexander
A
Triolo
Gianluca
G
Djeliova
Vera
V
Radulescu
Sorina
S
Vindigni
Alessandro
A
Riva
Silvano
S
Falaschi
Arturo
A
D001483:Base Sequence; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D034921:Lamin Type B / Q000378:metabolism; D007834:Lasers; D064126:Minichromosome Maintenance Complex Component 3; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D051738:Origin Recognition Complex; D018741:Replication Origin; D016196:S Phase; D013997:Time Factors; D014466:Ultraviolet Rays / Q000009:adverse effects*
C103706:CDC6 protein, human; D018797:Cell Cycle Proteins; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D034921:Lamin Type B; C066470:MCM3 protein, human; D009687:Nuclear Proteins; C497223:ORC1 protein, human; C497224:ORC2 protein, human; D051738:Origin Recognition Complex; D004247:DNA; D064126:Minichromosome Maintenance Complex Component 3