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PMID: 20347844 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

RCC1 uses a conformationally diverse loop region to interact with the nucleosome: a model for the RCC1-nucleosome complex.

Journal of molecular biology ·Vol. 398 ·No. 4 ·2010-05-14 ·Pages 518-29

England JR, Huang J, Jennings MJ, Makde RD, Tan S

Abstract

The binding of RCC1 (regulator of chromosome condensation 1) to chromatin is critical for cellular processes such as mitosis, nucleocytoplasmic transport, and nuclear envelope formation because RCC1 recruits the small GTPase Ran (Ras-related nuclear protein) to chromatin and sets up a Ran-GTP gradient around the chromosomes. However, the molecular mechanism by which RCC1 binds to nucleosomes, the repeating unit of chromatin, is not known. We have used biochemical approaches to test structural models for how the RCC1 beta-propeller protein could bind to the nucleosome. In contrast to the prevailing model, RCC1 does not appear to use the beta-propeller face opposite to its Ran-binding face to interact with nucleosomes. Instead, we find that RCC1 uses a conformationally flexible loop region we have termed the switchback loop in addition to its N-terminal tail to bind to the nucleosome. The juxtaposition of the RCC1 switchback loop to its Ran binding surface suggests a novel mechanism for how nucleosome-bound RCC1 recruits Ran to chromatin. Furthermore, this model accounts for previously unexplained observations for how Ran can interact with the nucleosome both dependent and independent of RCC1 and how binding of the nucleosome can enhance RCC1's Ran nucleotide exchange activity.

MeSH Terms
Amino Acid Substitution/genetics Cell Cycle Proteins/metabolism Guanine Nucleotide Exchange Factors/metabolism Models, Molecular Mutagenesis, Site-Directed Nuclear Proteins/metabolism Nucleosomes/metabolism Protein Binding Protein Conformation Protein Interaction Mapping Protein Structure, Tertiary
Chemicals
Cell Cycle Proteins Guanine Nucleotide Exchange Factors Nuclear Proteins Nucleosomes RCC1 protein, human
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
England Joseph R
Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, 108 Althouse Laboratory, The Pennsylvania State University, University Park, PA 16802-1014, USA.
Huang Jiehuan
Jennings Matthew J
Makde Ravindra D
Tan Song
References (36)
36 references, click to expand
  1. Transport into and out of the nucleus.
    Microbiol Mol Biol Rev. 2001 Dec;65(4):570-94, table of contents PMID: 11729264
  2. Comparison of affinity tags for protein purification.
    Protein Expr Purif. 2005 May;41(1):98-105 PMID: 15802226
  3. Nucleocytoplasmic transport: Ran, beta and beyond.
    Trends Cell Biol. 2001 Dec;11(12):497-503 PMID: 11719056
  4. Mechanisms of receptor-mediated nuclear import and nuclear export.
    Traffic. 2005 Mar;6(3):187-98 PMID: 15702987
  5. N-terminal alpha-methylation of RCC1 is necessary for stable chromatin association and normal mitosis.
    Nat Cell Biol. 2007 May;9(5):596-603 PMID: 17435751
  6. Nucleocytoplasmic transport of proteins.
    Biochemistry (Mosc). 2007 Dec;72(13):1439-57 PMID: 18282135
  7. The nucleosome: from genomic organization to genomic regulation.
    Cell. 2004 Jan 23;116(2):259-72 PMID: 14744436
  8. A modular polycistronic expression system for overexpressing protein complexes in Escherichia coli.
    Protein Expr Purif. 2001 Feb;21(1):224-34 PMID: 11162410
  9. WDR5 interacts with mixed lineage leukemia (MLL) protein via the histone H3-binding pocket.
    J Biol Chem. 2008 Dec 12;283(50):35258-64 PMID: 18840606
  10. Ran binds to chromatin by two distinct mechanisms.
    Curr Biol. 2002 Jul 9;12(13):1151-6 PMID: 12121625
  11. New DNA sequence rules for high affinity binding to histone octamer and sequence-directed nucleosome positioning.
    J Mol Biol. 1998 Feb 13;276(1):19-42 PMID: 9514715
  12. DNA-binding domain of RCC1 protein is not essential for coupling mitosis with DNA replication.
    J Cell Sci. 1992 Jul;102 ( Pt 3):393-400 PMID: 1506422
  13. Molecular mechanism of the nuclear protein import cycle.
    Nat Rev Mol Cell Biol. 2007 Mar;8(3):195-208 PMID: 17287812
  14. Molecular recognition of histone H3 by the WD40 protein WDR5.
    Nat Struct Mol Biol. 2006 Aug;13(8):698-703 PMID: 16829960
  15. The role of chromatin during transcription.
    Cell. 2007 Feb 23;128(4):707-19 PMID: 17320508
  16. Sequence motifs and free energies of selected natural and non-natural nucleosome positioning DNA sequences.
    J Mol Biol. 1999 Apr 30;288(2):213-29 PMID: 10329138
  17. Structural basis for guanine nucleotide exchange on Ran by the regulator of chromosome condensation (RCC1).
    Cell. 2001 Apr 20;105(2):245-55 PMID: 11336674
  18. Chromatin docking and exchange activity enhancement of RCC1 by histones H2A and H2B.
    Science. 2001 May 25;292(5521):1540-3 PMID: 11375490
  19. Primers on chromatin.
    Nat Struct Mol Biol. 2007 Nov;14(11):1110-5 PMID: 17984971
  20. Structural and molecular characterization of a preferred protein interaction surface on G protein beta gamma subunits.
    Biochemistry. 2005 Aug 9;44(31):10593-604 PMID: 16060668
  21. Keeping G proteins at bay: a complex between G protein-coupled receptor kinase 2 and Gbetagamma.
    Science. 2003 May 23;300(5623):1256-62 PMID: 12764189
  22. The pST44 polycistronic expression system for producing protein complexes in Escherichia coli.
    Protein Expr Purif. 2005 Apr;40(2):385-95 PMID: 15766881
  23. Chromatin-modifying enzymes as therapeutic targets--Part 1.
    Expert Opin Ther Targets. 2008 Oct;12(10):1301-12 PMID: 18781828
  24. Expression and purification of recombinant histones and nucleosome reconstitution.
    Methods Mol Biol. 1999;119:1-16 PMID: 10804500
  25. Structural basis for molecular recognition and presentation of histone H3 by WDR5.
    EMBO J. 2006 Sep 20;25(18):4245-52 PMID: 16946699
  26. The 1.7 A crystal structure of the regulator of chromosome condensation (RCC1) reveals a seven-bladed propeller.
    Nature. 1998 Mar 5;392(6671):97-101 PMID: 9510255
  27. Concentration of Ran on chromatin induces decondensation, nuclear envelope formation and nuclear pore complex assembly.
    Eur J Cell Biol. 2002 Nov;81(11):623-33 PMID: 12494999
  28. Histone H3 recognition and presentation by the WDR5 module of the MLL1 complex.
    Nat Struct Mol Biol. 2006 Aug;13(8):704-12 PMID: 16829959
  29. Structural basis for phosphodependent substrate selection and orientation by the SCFCdc4 ubiquitin ligase.
    Cell. 2003 Jan 24;112(2):243-56 PMID: 12553912
  30. Regulation of chromatin binding by a conformational switch in the tail of the Ran exchange factor RCC1.
    J Cell Biol. 2008 Sep 8;182(5):827-36 PMID: 18762580
  31. Ran at a glance.
    J Cell Sci. 2006 Sep 1;119(Pt 17):3481-4 PMID: 16931595
  32. Snapshot of activated G proteins at the membrane: the Galphaq-GRK2-Gbetagamma complex.
    Science. 2005 Dec 9;310(5754):1686-90 PMID: 16339447
  33. The RCC1 protein, a regulator for the onset of chromosome condensation locates in the nucleus and binds to DNA.
    J Cell Biol. 1989 Oct;109(4 Pt 1):1389-97 PMID: 2677018
  34. New nomenclature for chromatin-modifying enzymes.
    Cell. 2007 Nov 16;131(4):633-6 PMID: 18022353
  35. Structure of WDR5 bound to mixed lineage leukemia protein-1 peptide.
    J Biol Chem. 2008 Nov 21;283(47):32158-61 PMID: 18829459
  36. The nucleosomal surface as a docking station for Kaposi's sarcoma herpesvirus LANA.
    Science. 2006 Feb 10;311(5762):856-61 PMID: 16469929
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
1089-8638
Published
2010-05-14
Epub
2010-00-27
Pages
518-29
Language
English
Region
England
NLM ID
2985088R
PMCID
PMC2895563
Subset
IM
Grants
NIGMS NIH HHS · R01 GM088236 · United States
NIGMS NIH HHS · R01 GM088236-01 · United States
NIGMS NIH HHS · GM088236 · United States
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