Siddiqui, K., Stillman, B. (November 2007) ATP dependent assembly of the human origin recognition complex. Journal of Biological Chemistry, 282 (44). pp. 32370-32383. ISSN 0021-9258
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Abstract
The Origin Recognition Complex (ORC) was initially discovered in budding yeast extracts as a protein complex that binds with high affinity to Autonomously Replicating Sequences (ARS) in an ATP dependent manner. We have cloned and expressed the human homologs of the ORC subunits as recombinant proteins. In contrast to other eukaryotic initiators examined thus far, assembly of human ORC in vitro is dependent on ATP binding. Mutations in the ATP binding sites of Orc4 or Orc5 impair complex assembly, whereas Orc1 ATP binding is not required. Immunofluorescence staining of human cells with anti-Orc3 antibodies demonstrate cell cycle-dependent association with a nuclear structure. Immunoprecipitation experiments show that ORC disassembles as cells progress through S phase. The Orc6 protein binds directly to the Orc3 subunit and interacts as part of ORC in vivo. These data suggest that the assembly and disassembly of ORC in human cells is uniquely regulated and may contribute to restricting DNA replication to once in every cell division cycle.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | EUKARYOTIC DNA-REPLICATION WALKER-A MOTIF CONFORMATIONAL-CHANGES ESCHERICHIA-COLI CELL-CYCLE IN-VITRO HEXAMERIC HELICASE T-ANTIGEN CHROMATIN PROTEINS |
| Subjects: | organism description > animal > mammal > primates > hominids > human bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs > Stillman lab |
| Highlight: | Stillman, Bruce W. |
| Depositing User: | CSHL Librarian |
| Date: | November 2007 |
| Date Deposited: | 07 Nov 2011 18:36 |
| Last Modified: | 20 Jun 2017 16:42 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/23139 |
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