Localization of proteins bound to a replication origin of human DNA along the cell cycle

Gulnara Abdurashidova; Miltcho B. Danailov; Alexander Ochem; Gianluca Triolo; Vera Djeliova; Sorina Radulescu; Alessandro Vindigni; Silvano Riva; Arturo Falaschi

doi:10.1093/emboj/cdg404

Localization of proteins bound to a replication origin of human DNA along the cell cycle

Gulnara Abdurashidova, Miltcho B. Danailov, Alexander Ochem, Gianluca Triolo, Vera Djeliova, Sorina Radulescu, Alessandro Vindigni, Silvano Riva, Arturo Falaschi

Research output: Contribution to journal › Article › peer-review

65 Scopus citations

Abstract

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 G₁, 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.

Original language	English
Pages (from-to)	4294-4303
Number of pages	10
Journal	EMBO Journal
Volume	22
Issue number	16
DOIs	https://doi.org/10.1093/emboj/cdg404
State	Published - Aug 15 2003

Keywords

Lamin B2 origin
ORC proteins
Origin of replication
UV cross-linking

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10.1093/emboj/cdg404

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title = "Localization of proteins bound to a replication origin of human DNA along the cell cycle",

abstract = "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 G1, 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.",

keywords = "Lamin B2 origin, ORC proteins, Origin of replication, UV cross-linking",

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AU - Abdurashidova, Gulnara

AU - Danailov, Miltcho B.

AU - Ochem, Alexander

AU - Triolo, Gianluca

AU - Djeliova, Vera

AU - Radulescu, Sorina

AU - Vindigni, Alessandro

AU - Riva, Silvano

AU - Falaschi, Arturo

PY - 2003/8/15

Y1 - 2003/8/15

N2 - 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 G1, 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.

AB - 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 G1, 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.

KW - Lamin B2 origin

KW - ORC proteins

KW - Origin of replication

KW - UV cross-linking

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JF - EMBO Journal

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Localization of proteins bound to a replication origin of human DNA along the cell cycle

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Keywords

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