title: RNA binding and chaperone activity of the E. coli cold-shock protein CspA.
abstract: Ensuring the correct folding of RNA molecules in the cell is of major importance for a large variety of biological functions. Therefore, chaperone proteins that assist RNA in adopting their functionally active states are abundant in all living organisms. An important feature of RNA chaperone proteins is that they do not require an external energy source to perform their activity, and that they interact transiently and non-specifically with their RNA targets. So far, little is known about the mechanistic details of the RNA chaperone activity of these proteins. Prominent examples of RNA chaperones are bacterial cold shock proteins (Csp) that have been reported to bind single-stranded RNA and DNA. Here, we have used advanced NMR spectroscopy techniques to investigate at atomic resolution the RNA-melting activity of CspA, the major cold shock protein of Escherichia coli, upon binding to different RNA hairpins. Real-time NMR provides detailed information on the folding kinetics and folding pathways. Finally, comparison of wild-type CspA with single-point mutants and small peptides yields insights into the complementary roles of aromatic and positively charged amino-acid side chains for the RNA chaperone activity of the protein.
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Enrico
authors.initials: E
authors.lastname: Rennella
authors.affiliation: Department of Computational & Structural Biology, Max F. Perutz Laboratories, Campus, Vienna Biocenter 5, A-1030 Vienna, Austria.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Sára
authors.affiliation: Institute of Organic Chemistry, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Juen
authors.affiliation: Institute of Organic Chemistry, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Wunderlich
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Imbert
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Zsofia
authors.initials: Z
authors.lastname: Solyom
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Adrien
authors.initials: A
authors.lastname: Favier
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Isabel
authors.initials: I
authors.lastname: Ayala
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Katharina
authors.initials: K
authors.lastname: Weinhäupl
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Paul
authors.initials: P
authors.lastname: Schanda
authors.affiliation: Department of Computational & Structural Biology, Max F. Perutz Laboratories, Campus, Vienna Biocenter 5, A-1030 Vienna, Austria.
authors.forename: Robert
authors.initials: R
authors.lastname: Konrat
authors.affiliation: Institute of Organic Chemistry, University of Innsbruck, Innrain 80/82, 6020 Innsbruck, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Kreutz
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044 Grenoble Cedex 9, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.|Centre National de Recherche Scientifique (CNRS), Grenoble, France.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Brutscher
mesh_terms: D024322:Amino Acids, Aromatic / Q000737:chemistry; D057773:Cold Shock Proteins and Peptides / Q000737:chemistry* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011487:Protein Conformation; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D059370:RNA Folding
chemical_list: D024322:Amino Acids, Aromatic; D057773:Cold Shock Proteins and Peptides; C498192:CspA protein, E coli; D029968:Escherichia coli Proteins; D012313:RNA