Sequence-independent DNA Binding and Replication Initiation by the Human Origin Recognition Complex

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2003 Aug 5

PMID 12897055

Citations 171

Authors

Sanjay Vashee

Christin Cvetic

Wenyan Lu

Pamela Simancek

Thomas J Kelly

Johannes C Walter

Affiliations

Soon will be listed here.

Abstract

We report that a highly purified human origin recognition complex (HsORC) has intrinsic DNA-binding activity, and that this activity is modestly stimulated by ATP. HsORC binds preferentially to synthetic AT-rich polydeoxynucleotides, but does not effectively discriminate between natural DNA fragments that contain known human origins and control fragments. The complex fully restores DNA replication to ORC-depleted Xenopus egg extracts, providing strong evidence for its initiator function. Strikingly, HsORC stimulates initiation from any DNA sequence, and it does not preferentially replicate DNA containing human origin sequences. These data provide a biochemical explanation for the observation that in metazoans, initiation of DNA replication often occurs in a seemingly random pattern, and they have important implications for the nature of human origins of DNA replication.

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References

Ina S, Sasaki T, Yokota Y, Shinomiya T . A broad replication origin of Drosophila melanogaster, oriDalpha, consists of AT-rich multiple discrete initiation sites. Chromosoma. 2001; 109(8):551-64. DOI: 10.1007/s004120000112. View

Vashee S, Simancek P, Challberg M, Kelly T . Assembly of the human origin recognition complex. J Biol Chem. 2001; 276(28):26666-73. DOI: 10.1074/jbc.M102493200. View

Dhar S, Delmolino L, Dutta A . Architecture of the human origin recognition complex. J Biol Chem. 2001; 276(31):29067-71. DOI: 10.1074/jbc.M103078200. View

Okuno Y, McNairn A, den Elzen N, Pines J, Gilbert D . Stability, chromatin association and functional activity of mammalian pre-replication complex proteins during the cell cycle. EMBO J. 2001; 20(15):4263-77. PMC: 149150. DOI: 10.1093/emboj/20.15.4263. View

Schepers A, Ritzi M, Bousset K, Kremmer E, Yates J, Harwood J . Human origin recognition complex binds to the region of the latent origin of DNA replication of Epstein-Barr virus. EMBO J. 2001; 20(16):4588-602. PMC: 125560. DOI: 10.1093/emboj/20.16.4588. View

Dhar S, Yoshida K, Machida Y, Khaira P, Chaudhuri B, Wohlschlegel J . Replication from oriP of Epstein-Barr virus requires human ORC and is inhibited by geminin. Cell. 2001; 106(3):287-96. DOI: 10.1016/s0092-8674(01)00458-5. View

Romanowski P, Madine M, Rowles A, J Blow J, Laskey R . The Xenopus origin recognition complex is essential for DNA replication and MCM binding to chromatin. Curr Biol. 1996; 6(11):1416-25. DOI: 10.1016/s0960-9822(96)00746-4. View

Walter J, Newport J . Regulation of replicon size in Xenopus egg extracts. Science. 1997; 275(5302):993-5. DOI: 10.1126/science.275.5302.993. View

Royzman I, Austin R, Bosco G, Bell S . ORC localization in Drosophila follicle cells and the effects of mutations in dE2F and dDP. Genes Dev. 1999; 13(7):827-40. PMC: 316602. DOI: 10.1101/gad.13.7.827. View

10.

Rowles A, Tada S, J Blow J . Changes in association of the Xenopus origin recognition complex with chromatin on licensing of replication origins. J Cell Sci. 1999; 112 ( Pt 12):2011-8. PMC: 3605702. DOI: 10.1242/jcs.112.12.2011. View

11.

Okuno Y, Satoh H, Sekiguchi M, Masukata H . Clustered adenine/thymine stretches are essential for function of a fission yeast replication origin. Mol Cell Biol. 1999; 19(10):6699-709. PMC: 84658. DOI: 10.1128/MCB.19.10.6699. View

12.

Antequera F, Bird A . CpG islands as genomic footprints of promoters that are associated with replication origins. Curr Biol. 1999; 9(17):R661-7. DOI: 10.1016/s0960-9822(99)80418-7. View

13.

Chaudhuri B, Xu H, Todorov I, Dutta A, Yates J . Human DNA replication initiation factors, ORC and MCM, associate with oriP of Epstein-Barr virus. Proc Natl Acad Sci U S A. 2001; 98(18):10085-9. PMC: 56919. DOI: 10.1073/pnas.181347998. View

14.

Gilbert D . Making sense of eukaryotic DNA replication origins. Science. 2001; 294(5540):96-100. PMC: 1255916. DOI: 10.1126/science.1061724. View

15.

Chesnokov I, Remus D, Botchan M . Functional analysis of mutant and wild-type Drosophila origin recognition complex. Proc Natl Acad Sci U S A. 2001; 98(21):11997-2002. PMC: 59756. DOI: 10.1073/pnas.211342798. View

16.

Takahashi T, Masukata H . Interaction of fission yeast ORC with essential adenine/thymine stretches in replication origins. Genes Cells. 2001; 6(10):837-49. DOI: 10.1046/j.1365-2443.2001.00468.x. View

17.

Kong D, DePamphilis M . Site-specific DNA binding of the Schizosaccharomyces pombe origin recognition complex is determined by the Orc4 subunit. Mol Cell Biol. 2001; 21(23):8095-103. PMC: 99975. DOI: 10.1128/MCB.21.23.8095-8103.2001. View

18.

Lee J, Moon K, Jiang Y, Hurwitz J . The Schizosaccharomyces pombe origin recognition complex interacts with multiple AT-rich regions of the replication origin DNA by means of the AT-hook domains of the spOrc4 protein. Proc Natl Acad Sci U S A. 2001; 98(24):13589-94. PMC: 61085. DOI: 10.1073/pnas.251530398. View

19.

Lee S, Richardson C . Essential lysine residues in the RNA polymerase domain of the gene 4 primase-helicase of bacteriophage T7. J Biol Chem. 2001; 276(52):49419-26. DOI: 10.1074/jbc.M108443200. View

20.

Ladenburger E, Keller C, Knippers R . Identification of a binding region for human origin recognition complex proteins 1 and 2 that coincides with an origin of DNA replication. Mol Cell Biol. 2002; 22(4):1036-48. PMC: 134626. DOI: 10.1128/MCB.22.4.1036-1048.2002. View