Home LiteratureArticle Details
PMID: 11809796 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of a binding region for human origin recognition complex proteins 1 and 2 that coincides with an origin of DNA replication.

Molecular and cellular biology ·Vol. 22 ·No. 4 ·2002-02-00 ·Pages 1036-48

Ladenburger EM, Keller C, Knippers R

Abstract

We investigated the binding regions of components of the origin recognition complex (ORC) in the human genome. For this purpose, we performed chromatin immunoprecipitation assays with antibodies against human Orc1 and Orc2 proteins. We identified a binding region for human Orc proteins 1 and 2 in a <1-kbp segment between two divergently transcribed human genes. The region is characterized by CpG tracts and a central sequence rich in AT base pairs. Both, Orc1 and Orc2 proteins are found at the intergenic region in the G(1) phase, but S-phase chromatin contains only Orc2 protein, supporting the notion that Orc1p dissociates from its binding site in the S phase. Sequences corresponding to the intergenic region are highly abundant in a fraction of nascent DNA strands, strongly suggesting that this region not only harbors the binding sites for Orc1 protein and Orc2 protein but also serves as an origin of bidirectional DNA replication.

MeSH Terms
Base Sequence Binding Sites Cell Cycle Proteins/genetics Chromatin/metabolism Cross-Linking Reagents/chemistry DNA/chemistry,metabolism DNA Replication DNA-Activated Protein Kinase DNA-Binding Proteins/chemistry,metabolism Formaldehyde/chemistry HeLa Cells Humans Lamins Micrococcal Nuclease/metabolism Minichromosome Maintenance Complex Component 4 Molecular Sequence Data Nuclear Proteins/genetics,metabolism Origin Recognition Complex Polymerase Chain Reaction Precipitin Tests Promoter Regions, Genetic Protein Binding Protein Serine-Threonine Kinases/genetics Replication Origin S Phase
Chemicals
Cell Cycle Proteins Chromatin Cross-Linking Reagents DNA-Binding Proteins Lamins Nuclear Proteins ORC1 protein, human ORC2 protein, human Origin Recognition Complex Formaldehyde DNA DNA-Activated Protein Kinase PRKDC protein, human Protein Serine-Threonine Kinases Micrococcal Nuclease MCM4 protein, human Minichromosome Maintenance Complex Component 4
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ladenburger Eva-Maria
Department of Biology, Universität Konstanz, D-78464 Konstanz, Germany. Eva.Ladenburger@uni-konstanz.de
Keller Christian
Knippers Rolf
References (62)
62 references, click to expand
  1. Start sites of bidirectional DNA synthesis at the human lamin B2 origin.
    Science. 2000 Mar 17;287(5460):2023-6 PMID: 10720330
  2. Mapping chromosomal proteins in vivo by formaldehyde-crosslinked-chromatin immunoprecipitation.
    Trends Biochem Sci. 2000 Mar;25(3):99-104 PMID: 10694875
  3. Selective instability of Orc1 protein accounts for the absence of functional origin recognition complexes during the M-G(1) transition in mammals.
    EMBO J. 2000 Jun 1;19(11):2728-38 PMID: 10835370
  4. Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis.
    Mol Cell Biol. 2000 Nov;20(22):8602-12 PMID: 11046155
  5. SV40 large T antigen functions in DNA replication and transformation.
    Adv Virus Res. 2000;55:75-134 PMID: 11050941
  6. Identification and characterization of the human ORC6 homolog.
    J Biol Chem. 2000 Nov 10;275(45):34983-8 PMID: 10945994
  7. Genome-wide location and function of DNA binding proteins.
    Science. 2000 Dec 22;290(5500):2306-9 PMID: 11125145
  8. Where it all starts: eukaryotic origins of DNA replication.
    J Cell Sci. 2001 Feb;114(Pt 4):643-51 PMID: 11171369
  9. Replication origins in yeast versus metazoa: separation of the haves and the have nots.
    Curr Opin Genet Dev. 1998 Apr;8(2):194-9 PMID: 9610410
  10. Nucleosome assembly activity and intracellular localization of human CAF-1 changes during the cell division cycle.
    J Biol Chem. 1998 Jun 12;273(24):15279-86 PMID: 9614144
  11. Nucleotide-dependent prereplicative complex assembly by Cdc6p, a homolog of eukaryotic and prokaryotic clamp-loaders.
    Mol Cell. 1998 Jul;2(1):23-32 PMID: 9702188
  12. Human minichromosome maintenance proteins and human origin recognition complex 2 protein on chromatin.
    J Biol Chem. 1998 Sep 18;273(38):24543-9 PMID: 9733749
  13. ORC5L, a new member of the human origin recognition complex, is deleted in uterine leiomyomas and malignant myeloid diseases.
    J Biol Chem. 1998 Oct 16;273(42):27137-45 PMID: 9765232
  14. The Orc4p and Orc5p subunits of the Xenopus and human origin recognition complex are related to Orc1p and Cdc6p.
    J Biol Chem. 1998 Dec 4;273(49):32421-9 PMID: 9829972
  15. The Cdc6p nucleotide-binding motif is required for loading mcm proteins onto chromatin.
    Proc Natl Acad Sci U S A. 1999 Jan 19;96(2):441-6 PMID: 9892652
  16. Replication origins in metazoan chromosomes: fact or fiction?
    Bioessays. 1999 Jan;21(1):5-16 PMID: 10070250
  17. Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F.
    Oncogene. 1999 Apr 8;18(14):2299-309 PMID: 10327050
  18. Multistep regulation of DNA replication by Cdk phosphorylation of HsCdc6.
    Proc Natl Acad Sci U S A. 1999 May 25;96(11):6193-8 PMID: 10339564
  19. Changes in association of the Xenopus origin recognition complex with chromatin on licensing of replication origins.
    J Cell Sci. 1999 Jun;112 ( Pt 12):2011-8 PMID: 10341218
  20. Chromatin immunoprecipitation assays in acetylation mapping of higher eukaryotes.
    Methods Enzymol. 1999;304:533-47 PMID: 10372380
  21. Plaque formation and isolation of pure lines with poliomyelitis viruses.
    J Exp Med. 1954 Feb;99(2):167-82 PMID: 13130792
  22. The human origin recognition complex protein 1 dissociates from chromatin during S phase in HeLa cells.
    J Biol Chem. 2001 Mar 2;276(9):6337-42 PMID: 11102449
  23. Assembly of the human origin recognition complex.
    J Biol Chem. 2001 Jul 13;276(28):26666-73 PMID: 11323433
  24. Architecture of the human origin recognition complex.
    J Biol Chem. 2001 Aug 3;276(31):29067-71 PMID: 11395502
  25. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  26. Eucaryotic DNA: organization of the genome for replication.
    Cell. 1978 Oct;15(2):317-25 PMID: 719745
  27. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  28. Purification of a yeast protein that binds to origins of DNA replication and a transcriptional silencer.
    Proc Natl Acad Sci U S A. 1988 Apr;85(7):2120-4 PMID: 3281162
  29. A yeast chromosomal origin of DNA replication defined by multiple functional elements.
    Science. 1992 Feb 14;255(5046):817-23 PMID: 1536007
  30. ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex.
    Nature. 1992 May 14;357(6374):128-34 PMID: 1579162
  31. The gene for a novel human lamin maps at a highly transcribed locus of chromosome 19 which replicates at the onset of S-phase.
    Mol Cell Biol. 1992 Aug;12(8):3499-506 PMID: 1630457
  32. A human DNA replication origin: localization and transcriptional characterization.
    Chromosoma. 1992;102(1 Suppl):S24-31 PMID: 1291239
  33. Eukaryotic DNA replication: anatomy of an origin.
    Annu Rev Biochem. 1993;62:29-63 PMID: 8352592
  34. Number of CpG islands and genes in human and mouse.
    Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11995-9 PMID: 7505451
  35. Fine mapping of a replication origin of human DNA.
    Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7119-23 PMID: 8041756
  36. Two steps in the assembly of complexes at yeast replication origins in vivo.
    Cell. 1994 Jul 29;78(2):303-16 PMID: 8044842
  37. The origin recognition complex interacts with a bipartite DNA binding site within yeast replicators.
    Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):2224-8 PMID: 7892251
  38. Purification of an MCM-containing complex as a component of the DNA replication licensing system.
    Nature. 1995 Jun 1;375(6530):418-21 PMID: 7760937
  39. MCM3 complex required for cell cycle regulation of DNA replication in vertebrate cells.
    Nature. 1995 Jun 1;375(6530):421-4 PMID: 7760938
  40. Initiation complex assembly at budding yeast replication origins begins with the recognition of a bipartite sequence by limiting amounts of the initiator, ORC.
    EMBO J. 1995 Jun 1;14(11):2631-41 PMID: 7781615
  41. Expression, phosphorylation and nuclear localization of the human P1 protein, a homologue of the yeast Mcm 3 replication protein.
    J Cell Sci. 1995 Apr;108 ( Pt 4):1381-9 PMID: 7615659
  42. Conserved initiator proteins in eukaryotes.
    Science. 1995 Dec 8;270(5242):1667-71 PMID: 7502077
  43. The Xenopus Cdc6 protein is essential for the initiation of a single round of DNA replication in cell-free extracts.
    Cell. 1996 Oct 4;87(1):53-63 PMID: 8858148
  44. Interaction between the origin recognition complex and the replication licensing system in Xenopus.
    Cell. 1996 Oct 18;87(2):287-96 PMID: 8861912
  45. The Xenopus origin recognition complex is essential for DNA replication and MCM binding to chromatin.
    Curr Biol. 1996 Nov 1;6(11):1416-25 PMID: 8939603
  46. A role for Cdk2 kinase in negatively regulating DNA replication during S phase of the cell cycle.
    J Cell Biol. 1997 Apr 7;137(1):183-92 PMID: 9105046
  47. The scaffold/matrix attachment region binding protein hnRNP-U (SAF-A) is directly bound to chromosomal DNA in vivo: a chemical cross-linking study.
    Biochemistry. 1997 Jul 8;36(27):8276-83 PMID: 9204873
  48. Expression, nuclear localization and interactions of human MCM/P1 proteins.
    Biochem Biophys Res Commun. 1997 Jul 9;236(1):118-25 PMID: 9223437
  49. MCM4 and PRKDC, human genes encoding proteins MCM4 and DNA-PKcs, are close neighbours located on chromosome 8q12-->q13.
    Cytogenet Cell Genet. 1997;77(3-4):268-70 PMID: 9284934
  50. Components and dynamics of DNA replication complexes in S. cerevisiae: redistribution of MCM proteins and Cdc45p during S phase.
    Cell. 1997 Oct 3;91(1):59-69 PMID: 9335335
  51. Identification of HsORC4, a member of the human origin of replication recognition complex.
    J Biol Chem. 1997 Nov 7;272(45):28247-51 PMID: 9353276
  52. Architecture of the yeast origin recognition complex bound to origins of DNA replication.
    Mol Cell Biol. 1997 Dec;17(12):7159-68 PMID: 9372948
  53. Discrete start sites for DNA synthesis in the yeast ARS1 origin.
    Science. 1998 Jan 2;279(5347):95-8 PMID: 9417033
  54. Mapping replication origins by quantifying relative abundance of nascent DNA strands using competitive polymerase chain reaction.
    Methods. 1997 Nov;13(3):301-12 PMID: 9441856
  55. The promoters for human DNA-PKcs (PRKDC) and MCM4: divergently transcribed genes located at chromosome 8 band q11.
    Genomics. 1998 Jan 1;47(1):71-83 PMID: 9465298
  56. Initiation of DNA replication at CpG islands in mammalian chromosomes.
    EMBO J. 1998 Apr 15;17(8):2426-35 PMID: 9545253
  57. Human CDC6/Cdc18 associates with Orc1 and cyclin-cdk and is selectively eliminated from the nucleus at the onset of S phase.
    Mol Cell Biol. 1998 May;18(5):2758-67 PMID: 9566895
  58. ORC binding, gene amplification, and the nature of metazoan replication origins.
    Genes Dev. 1999 Oct 15;13(20):2619-23 PMID: 10541547
  59. The metazoan origin recognition complex.
    Front Biosci. 1999 Dec 1;4:D805-15 PMID: 10577394
  60. Selection and mapping of replication origins from a 500-kb region of the human X chromosome and their relationship to gene expression.
    Genomics. 1999 Nov 15;62(1):11-20 PMID: 10585763
  61. The replication capacity of intact mammalian nuclei in Xenopus egg extracts declines with quiescence, but the residual DNA synthesis is independent of Xenopus MCM proteins.
    J Cell Sci. 2000 Feb;113 ( Pt 4):683-95 PMID: 10652261
  62. XCDT1 is required for the assembly of pre-replicative complexes in Xenopus laevis.
    Nature. 2000 Apr 6;404(6778):622-5 PMID: 10766247
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.22.4.1036-1048.2002
Email
eva.ladenburger@uni-konstanz.de
Published
2002-02-00
Pages
1036-48
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC134626
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.22.4.1036-1048.2002 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2