Single-stranded DNA bound to bacterial cold-shock proteins: preliminary crystallographic and Raman analysis

• Published in: Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2004-04, Vol.60 (4), p.755-757
• Main Authors: Bienert, Ralf, Zeeb, Markus, Dostál, Lubomir, Feske, Anette, Magg, Christine, Max, Klaas, Welfle, Heinz, Balbach, Jochen, Heinemann, Udo
• Format: Article
• Language: English
• Subjects: Bacillus - chemistry; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Cloning, Molecular; Crystallization; Crystallography, X-Ray; DNA, Single-Stranded - chemistry; DNA, Single-Stranded - metabolism; Heat-Shock Proteins - chemistry; Heat-Shock Proteins - metabolism; major cold-shock proteins; OB fold; Oligodeoxyribonucleotides - chemistry; Oligodeoxyribonucleotides - metabolism; protein-DNA interaction; Raman spectroscopy; single-stranded DNA; Spectrum Analysis, Raman
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S0907444904002422

The cold‐shock response has been described for several bacterial species. It is characterized by distinct changes in intracellular protein patterns whereby a set of cold‐shock‐inducible proteins become abundant. The major cold‐shock proteins of Bacillus subtilis (Bs‐­CspB) and Bacillus caldolyticus (Bc‐Csp) are small oligonucleotide/oligosaccharide‐binding (OB) fold proteins that have been described as binding single‐stranded nucleic acids. Bs‐CspB (Mr = 7365) and Bc‐Csp (Mr = 7333) were crystallized in the presence of the deoxyhexanucleotide (dT)6. Crystals of (dT)6 with Bs‐CspB grew in the orthorhombic space group C2221, with unit‐cell parameters a = 49.0, b = 53.2, c = 77.0 Å. Crystals with Bc‐Csp grew in the primitive orthorhombic space group P21212, with unit‐cell parameters a = 74.3, b = 64.9, c = 31.2 Å. These crystals diffract to maximal resolutions of 1.78 and 1.29 Å, respectively. The presence of protein and DNA in the crystals was demonstrated by Raman spectroscopy.