Expression, crystallization and preliminary crystallographic analysis of RNA-binding protein Hfq (YmaH) from Bacillus subtilis in complex with an RNA aptamer.
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.
Baba
Seiki
S
Japan Synchrotron Radiation Research Institute/SPring-8, Japan.
Someya
Tatsuhiko
T
Kawai
Gota
G
Nakamura
Kouji
K
Kumasaka
Takashi
T
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*
D052157:Aptamers, Nucleotide; D035001:Host Factor 1 Protein
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