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PMID:20932478

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Citation

Shen, Z, Sathyan, KM, Geng, Y, Zheng, R, Chakraborty, A, Freeman, B, Wang, F, Prasanth, KV and Prasanth, SG (2010) A WD-repeat protein stabilizes ORC binding to chromatin. Mol. Cell 40:99-111

Abstract

Origin recognition complex (ORC) plays critical roles in the initiation of DNA replication and cell-cycle progression. In metazoans, ORC associates with origin DNA during G1 and with heterochromatin in postreplicated cells. However, what regulates the binding of ORC to chromatin is not understood. We have identified a highly conserved, leucine-rich repeats and WD40 repeat domain-containing protein 1 (LRWD1) or ORC-associated (ORCA) in human cells that interacts with ORC and modulates chromatin association of ORC. ORCA colocalizes with ORC and shows similar cell-cycle dynamics. We demonstrate that ORCA efficiently recruits ORC to chromatin. Depletion of ORCA in human primary cells and embryonic stem cells results in loss of ORC association to chromatin, concomitant reduction of MCM binding, and a subsequent accumulation in G1 phase. Our results suggest ORCA-mediated association of ORC to chromatin is critical to initiate preRC assembly in G1 and chromatin organization in post-G1 cells.

Links

PubMed Online version:10.1016/j.molcel.2010.09.021

Keywords

Adaptor Proteins, Signal Transducing/genetics; Adaptor Proteins, Signal Transducing/metabolism; Amino Acid Sequence; Animals; Binding Sites; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Chromatin Assembly and Disassembly; Conserved Sequence; Fluorescent Antibody Technique; Heterochromatin/metabolism; Humans; Immunoprecipitation; Microscopy, Video; Molecular Sequence Data; Mutation; Origin Recognition Complex/genetics; Origin Recognition Complex/metabolism; Protein Binding; Protein Interaction Domains and Motifs; RNA Interference; Recombinant Fusion Proteins/metabolism; Time Factors; Transfection

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:ORC5

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ORC4

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:GEMI

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UFC0

F

Seeded From UniProt

complete

HUMAN:ORC1

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ORC2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UFC0

F

Seeded From UniProt

complete

HUMAN:CDT1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9UFC0

F

Seeded From UniProt

complete

HUMAN:LRWD1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13416

F

Seeded From UniProt

complete

HUMAN:LRWD1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O75496

F

Seeded From UniProt

complete

HUMAN:LRWD1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9H211

F

Seeded From UniProt

complete

HUMAN:LRWD1

involved_in

GO:0006325: chromatin organization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:LRWD1

involved_in

GO:0006270: DNA replication initiation

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

HUMAN:LRWD1

located_in

GO:0005721: pericentric heterochromatin

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:LRWD1

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:LRWD1

enables

GO:0003682: chromatin binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:LRWD1

involved_in

GO:0071169: establishment of protein localization to chromatin

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:LRWD1

located_in

GO:0000781: chromosome, telomeric region

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ORC3

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:ORC2

part_of

GO:0005664: nuclear origin of replication recognition complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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