Studies with the human cohesin establishment factor, ChlR1: Association of ChlR1 with Ctf18-RFC and Fen1 Journal Article
| Authors: | Farina, A.; Shin, J. H.; Kim, D. H.; Bermudez, V. P.; Kelman, Z.; Seo, Y. S.; Hurwitz, J. |
| Article Title: | Studies with the human cohesin establishment factor, ChlR1: Association of ChlR1 with Ctf18-RFC and Fen1 |
| Abstract: | Human ChlR1 (hChlR1), a member of the DEAD/DEAH subfamily of helicases, was shown to interact with components of the cohesin complex and play a role in sister chromatid cohesion. In order to study the biochemical and biological properties of hChlR1, we purified the protein from 293 cells and demonstrated that hChlR1 possesses DNA-dependent ATPase and helicase activities. This helicase translocates on single-stranded DNA in the 5′ to 3′ direction in the presence of ATP and, to a lesser extent, dATP. Its unwinding activity requires a 5′-single-stranded region for helicase loading, since flush-ended duplex structures do not support unwinding. The helicase activity of hChlR1 is capable of displacing duplex regions up to 100 bp, which can be extended to 500 bp by RPA or the cohesion establishment factor, the Ctf18-RFC (replication factor C) complex. We show that hChlR1 interacts with the hCtf18-RFC complex, human proliferating cell nuclear antigen, and hFen1. The interactions between Fen1 and hChlR1 stimulate the flap endonuclease activity of Fen1. Selective depletion of either hChlR1 or Fen1 by targeted small interfering RNA treatment results in the precocious separation of sister chromatids. These findings are consistent with a role of hChlR1 in the establishment of sister chromatid cohesion and suggest that its action may contribute to lagging strand processing events important in cohesion. © 2008 by The American Society for Biochemistry and Molecular Biology, Inc. |
| Keywords: | controlled study; carrier protein; unclassified drug; protein localization; cohesin; cell cycle protein; cohesins; nonhistone protein; metabolism; cell cycle proteins; chromosomal proteins, non-histone; genes; biological model; models, biological; nuclear protein; protein depletion; protein; small interfering rna; protein binding; rna, small interfering; protein interaction; enzyme activity; hela cell; hela cells; physiology; chromatid; nuclear proteins; dead box protein; rna; dead-box rna helicases; chemistry; dna; protein purification; nucleic acids; carrier proteins; recombinant proteins; recombinant protein; helicase; proliferating cell nuclear antigens; replication factor c; organic acids; adenosine triphosphate; cycline; flap endonuclease; fen1 protein, human; single stranded dna; flap endonucleases; replication protein c; enzyme structure; oligonucleotide; dna helicases; duplex structures; endonuclease activities; helicase activities; complementary dna; oligonucleotides; dna, complementary; helicases; chromatids; proliferating cell nuclear antigen; adhesion; bio-chemicals; 293 cells; biological properties; lagging strands; lesser extents; replication factors; unwinding activities; chlr1; ctf18; deoxyadenosine triphosphate; fen1; chtf18 protein, human; ddx11 protein, human |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 283 |
| Issue: | 30 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2008-07-25 |
| Start Page: | 20925 |
| End Page: | 20936 |
| Language: | English |
| DOI: | 10.1074/jbc.M802696200 |
| PUBMED: | 18499658 |
| PROVIDER: | scopus |
| PMCID: | PMC2475708 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 12" - "Export Date: 17 November 2011" - "CODEN: JBCHA" - "Source: Scopus" |
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