title: Solution structure of the dimerization domain of ribosomal protein P2 provides insights for the structural organization of eukaryotic stalk.
abstract: The lateral stalk of ribosome is responsible for kingdom-specific binding of translation factors and activation of GTP hydrolysis that drives protein synthesis. In eukaryotes, the stalk is composed of acidic ribosomal proteins P0, P1 and P2 that constitute a pentameric P-complex in 1: 2: 2 ratio. We have determined the solution structure of the N-terminal dimerization domain of human P2 (NTD-P2), which provides insights into the structural organization of the eukaryotic stalk. Our structure revealed that eukaryotic stalk protein P2 forms a symmetric homodimer in solution, and is structurally distinct from the bacterial counterpart L12 homodimer. The two subunits of NTD-P2 form extensive hydrophobic interactions in the dimeric interface that buries 2400 A(2) of solvent accessible surface area. We have showed that P1 can dissociate P2 homodimer spontaneously to form a more stable P1/P2 1 : 1 heterodimer. By homology modelling, we identified three exposed polar residues on helix-3 of P2 are substituted by conserved hydrophobic residues in P1. Confirmed by mutagenesis, we showed that these residues on helix-3 of P1 are not involved in the dimerization of P1/P2, but instead play a vital role in anchoring P1/P2 heterodimer to P0. Based on our results, models of the eukaryotic stalk complex were proposed.
authors.affiliation: Department of Biochemistry, Centre for Protein Science and Crystallography, The Chinese University of Hong Kong, Hong Kong, China.
authors.forename: Ka-Ming
authors.initials: KM
authors.lastname: Lee
authors.forename: Conny Wing-Heng
authors.initials: CW
authors.lastname: Yu
authors.forename: Denise So-Bik
authors.initials: DS
authors.lastname: Chan
authors.forename: Teddy Yu-Hin
authors.initials: TY
authors.lastname: Chiu
authors.forename: Guang
authors.initials: G
authors.lastname: Zhu
authors.forename: Kong-Hung
authors.initials: KH
authors.lastname: Sze
authors.forename: Pang-Chui
authors.initials: PC
authors.lastname: Shaw
authors.forename: Kam-Bo
authors.initials: KB
authors.lastname: Wong
mesh_terms: D000595:Amino Acid Sequence; D019281:Dimerization; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010750:Phosphoproteins / Q000737:chemistry*; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000737:chemistry*; D012996:Solutions
chemical_list: D010750:Phosphoproteins; D012269:Ribosomal Proteins; D012996:Solutions; C031937:phosphoprotein P2, ribosomal; C035690:ribosomal phosphoprotein P1
references.citation: Nat Protoc. 2007;2(11):2728-33
references.pmid: 18007608
references.citation: J Comput Chem. 2004 Oct;25(13):1605-12
references.pmid: 15264254
references.citation: Biol Cell. 2003 May-Jun;95(3-4):179-93
references.pmid: 12867082
references.citation: J Biol Chem. 2004 Apr 23;279(17):17697-706
references.pmid: 14960595
references.citation: J Biomol NMR. 1995 Nov;6(3):277-93
references.pmid: 8520220
references.citation: J Biomol NMR. 1996 Dec;8(4):477-86
references.pmid: 9008363
references.citation: Genes Cells. 2007 Apr;12(4):501-10
references.pmid: 17397397
references.citation: J Biochem. 2008 Feb;143(2):169-77
references.pmid: 17984123
references.citation: J Biol Chem. 2002 Feb 8;277(6):3857-62
references.pmid: 11729183
references.citation: Methods Enzymol. 2001;339:71-90
references.pmid: 11462826
references.citation: Nucleic Acids Res. 2007;35(5):1660-72
references.pmid: 17308345
references.citation: Biochemistry. 2000 Aug 1;39(30):8929-34
references.pmid: 10913305
references.citation: J Biomol NMR. 1994 Sep;4(5):603-14
references.pmid: 22911360
references.citation: Curr Protein Pept Sci. 2002 Feb;3(1):93-106
references.pmid: 12370014
references.citation: Proc Natl Acad Sci U S A. 1987 Aug;84(16):5580-4
references.pmid: 3475694
references.citation: Biochem J. 2008 Aug 1;413(3):527-34
references.pmid: 18422483
references.citation: EMBO J. 2004 Mar 10;23(5):1008-19
references.pmid: 14976550
references.citation: Structure. 2008 Apr;16(4):535-48
references.pmid: 18400176
references.citation: J Biomol NMR. 1999 Mar;13(3):289-302
references.pmid: 10212987
references.citation: Annu Rev Biophys Biomol Struct. 2000;29:291-325
references.pmid: 10940251
references.citation: Nucleic Acids Res. 2002 Jun 15;30(12):2620-7
references.pmid: 12060678
references.citation: Nucleic Acids Res. 2009 Feb;37(2):602-10
references.pmid: 19073700
references.citation: J Mol Biol. 2008 Apr 25;378(2):375-86
references.pmid: 18358491
references.citation: Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
references.pmid: 9757107
references.citation: J Biol Chem. 1999 Sep 24;274(39):27578-82
references.pmid: 10488095
references.citation: EMBO J. 2000 Nov 15;19(22):6075-84
references.pmid: 11080154
references.citation: J Biol Chem. 2005 Nov 25;280(47):39193-9
references.pmid: 16188884
references.citation: Mol Microbiol. 2005 Oct;58(1):217-26
references.pmid: 16164560
references.citation: FEBS Lett. 1976 Apr 15;64(1):135-8
references.pmid: 773698
references.citation: EMBO J. 2000 Jan 17;19(2):174-86
references.pmid: 10637222
references.citation: Mol Microbiol. 2008 Dec;70(6):1441-52
references.pmid: 19019145
references.citation: J Biol Chem. 2010 Feb 12;285(7):4747-56
references.pmid: 20007716
references.citation: FEBS Lett. 1978 Sep 15;93(2):215-8
references.pmid: 361431
references.citation: Biochim Biophys Acta. 2000 Dec 11;1499(1-2):63-73
references.pmid: 11118639
references.citation: Biochemistry. 2010 Feb 9;49(5):924-33
references.pmid: 20058904
references.citation: Int J Biochem Cell Biol. 2000 Jul;32(7):737-46
references.pmid: 10856704
references.citation: EMBO J. 2000 Jun 1;19(11):2710-8
references.pmid: 10835368
references.citation: Biochemistry. 2003 Apr 1;42(12):3399-408
references.pmid: 12653543
references.citation: Mol Microbiol. 2006 Apr;60(2):386-400
references.pmid: 16573688
references.citation: Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1469-73
references.pmid: 1996347
references.citation: Prog Nucleic Acid Res Mol Biol. 1996;55:157-93
references.pmid: 8787610
references.citation: J Biol Chem. 2001 Jun 8;276(23):19762-9
references.pmid: 11274186
references.citation: Cell. 2005 Jul 1;121(7):991-1004
references.pmid: 15989950
references.citation: Biochemistry. 2007 Feb 20;46(7):1988-98
references.pmid: 17261029
references.citation: Biochemistry. 1988 Oct 18;27(21):8063-8
references.pmid: 3233195
references.citation: Int J Biochem Cell Biol. 2009 Jun;41(6):1315-22
references.pmid: 19084076