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Molecular and Cellular Biology
Article . 2012 . Peer-reviewed
License: ASM Journals Non-Commercial TDM
Data sources: Crossref
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Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation

Authors: Zhen, Shen; Arindam, Chakraborty; Ankur, Jain; Sumanprava, Giri; Taekjip, Ha; Kannanganattu V, Prasanth; Supriya G, Prasanth;

Dynamic Association of ORCA with Prereplicative Complex Components Regulates DNA Replication Initiation

Abstract

In eukaryotes, initiation of DNA replication requires the assembly of a multiprotein prereplicative complex (pre-RC) at the origins. We recently reported that a WD repeat-containing protein, origin recognition complex (ORC)-associated (ORCA/LRWD1), plays a crucial role in stabilizing ORC to chromatin. Here, we find that ORCA is required for the G(1)-to-S-phase transition in human cells. In addition to binding to ORC, ORCA associates with Cdt1 and its inhibitor, geminin. Single-molecule pulldown experiments demonstrate that each molecule of ORCA can bind to one molecule of ORC, one molecule of Cdt1, and two molecules of geminin. Further, ORCA directly interacts with the N terminus of Orc2, and the stability of ORCA is dependent on its association with Orc2. ORCA associates with Orc2 throughout the cell cycle, with Cdt1 during mitosis and G(1), and with geminin in post-G(1) cells. Overexpression of geminin results in the loss of interaction between ORCA and Cdt1, suggesting that increased levels of geminin in post-G(1) cells titrate Cdt1 away from ORCA. We propose that the dynamic association of ORCA with pre-RC components modulates the assembly of its interacting partners on chromatin and facilitates DNA replication initiation.

Keywords

Cell Nucleus, DNA Replication, Cell Cycle, Geminin, Origin Recognition Complex, Mitosis, Cell Cycle Proteins, G1 Phase Cell Cycle Checkpoints, Chromatin, DNA-Binding Proteins, Cell Line, Tumor, Sequestosome-1 Protein, Humans, Adaptor Proteins, Signal Transducing, HeLa Cells, Protein Binding

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    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
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    Top 10%
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    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
43
Top 10%
Top 10%
Top 10%
bronze