title: Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer.
abstract: The Hfq protein is a hexameric RNA-binding protein which regulates gene expression by binding to RNA under the influence of diverse environmental stresses. Its ring structure binds various types of RNA, including mRNA and sRNA. RNA-bound structures of Hfq from Escherichia coli and Staphylococcus aureus have been revealed to have poly(A) RNA at the distal site and U-rich RNA at the proximal site, respectively. Here, crystals of a complex of the Bacillus subtilis Hfq protein with an A/G-repeat 7-mer RNA (Hfq-RNA) that were prepared using the hanging-drop vapour-diffusion technique are reported. The type 1 Hfq-RNA crystals belonged to space group I422, with unit-cell parameters a = b = 123.70, c = 119.13 A, while the type 2 Hfq-RNA crystals belonged to space group F222, with unit-cell parameters a = 91.92, b = 92.50, c = 114.92 A. Diffraction data were collected to a resolution of 2.20 A from both crystal forms. The hexameric structure of the Hfq protein was clearly shown by self-rotation analysis.
authors.affiliation: Japan Synchrotron Radiation Research Institute/SPring-8, Japan.
authors.forename: Seiki
authors.initials: S
authors.lastname: Baba
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Someya
authors.forename: Gota
authors.initials: G
authors.lastname: Kawai
authors.forename: Kouji
authors.initials: K
authors.lastname: Nakamura
authors.forename: Takashi
authors.initials: T
authors.lastname: Kumasaka
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001412:Bacillus subtilis / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D035001:Host Factor 1 Protein / Q000737:chemistry*
chemical_list: D052157:Aptamers, Nucleotide; D035001:Host Factor 1 Protein
references.citation: Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1501-5
references.pmid: 10677490
references.citation: EMBO Rep. 2003 Mar;4(3):284-9
references.pmid: 12634847
references.citation: Nat Struct Mol Biol. 2004 Dec;11(12):1206-14
references.pmid: 15531892
references.citation: Mol Cell. 2002 Jan;9(1):11-22
references.pmid: 11804582
references.citation: RNA. 2007 Dec;13(12):2213-23
references.pmid: 17959927
references.citation: J Biol Chem. 1972 Feb 10;247(3):824-31
references.pmid: 4550762
references.citation: Nucleic Acids Res. 2003 Jul 15;31(14):4091-8
references.pmid: 12853626
references.citation: BMC Microbiol. 2007 Feb 09;7:10
references.pmid: 17291347
references.citation: Nucleic Acids Res. 2006 Jan 30;34(2):709-20
references.pmid: 16449205
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
references.pmid: 15299926
references.citation: Curr Opin Microbiol. 2007 Apr;10(2):125-33
references.pmid: 17395525
references.citation: Genes Dev. 2002 Jul 1;16(13):1696-706
references.pmid: 12101127
references.citation: Mol Microbiol. 2006 Oct;62(2):520-36
references.pmid: 17020585
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: EMBO J. 2002 Jul 1;21(13):3546-56
references.pmid: 12093755
references.citation: RNA. 2003 Jan;9(1):33-43
references.pmid: 12554874
references.citation: EMBO J. 2004 Jan 28;23(2):396-405
references.pmid: 14739933
references.citation: Acta Crystallogr D Biol Crystallogr. 2005 Feb;61(Pt 2):141-6
references.pmid: 15681864
references.citation: Mol Microbiol. 2004 Nov;54(4):905-20
references.pmid: 15522076
references.citation: Nucleic Acids Res. 2005 Mar 21;33(5):1678-89
references.pmid: 15781494
references.citation: Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12462-7
references.pmid: 9770508
references.citation: Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19292-7
references.pmid: 19889981
references.citation: FEBS J. 2005 Jan;272(2):454-63
references.pmid: 15654883
references.citation: J Mol Biol. 1968 Apr 28;33(2):491-7
references.pmid: 5700707
references.citation: Nucleic Acids Res. 2007;35(13):4331-46
references.pmid: 17576690