TY  - JOUR
A1  - Qureshi, Nusrat
A1  - Matzel, Tobias
A1  - Çetiner, Erhan Can
A1  - Schnieders, Robbin
A1  - Jonker, Hendrik R. A.
A1  - Schwalbe, Harald
A1  - Fürtig, Boris
T1  - NMR structure of the Vibrio vulnificus ribosomal protein S1 domains D3 and D4 provides insights into molecular recognition of single-stranded RNAs
T2  - Nucleic acids research
N2  - The ribosomal S1 protein (rS1) is indispensable for translation initiation in Gram-negative bacteria. rS1 is a multidomain protein that acts as an RNA chaperone and ensures that mRNAs can bind the ribosome in a single-stranded conformation, which could be related to fast recognition. Although many ribosome structures were solved in recent years, a high-resolution structure of a two-domain mRNA-binding competent rS1 construct is not yet available. Here, we present the NMR solution structure of the minimal mRNA-binding fragment of Vibrio Vulnificus rS1 containing the domains D3 and D4. Both domains are homologues and adapt an oligonucleotide-binding fold (OB fold) motif. NMR titration experiments reveal that recognition of miscellaneous mRNAs occurs via a continuous interaction surface to one side of these structurally linked domains. Using a novel paramagnetic relaxation enhancement (PRE) approach and exploring different spin-labeling positions within RNA, we were able to track the location and determine the orientation of the RNA in the rS1–D34 bound form. Our investigations show that paramagnetically labeled RNAs, spiked into unmodified RNA, can be used as a molecular ruler to provide structural information on protein-RNA complexes. The dynamic interaction occurs on a defined binding groove spanning both domains with identical β2-β3-β5 interfaces. Evidently, the 3′-ends of the cis-acting RNAs are positioned in the direction of the N-terminus of the rS1 protein, thus towards the 30S binding site and adopt a conformation required for translation initiation.
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/63304
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-633047
SN  - 1362-4962
N1  - German funding agency (DFG) in Collaborative Research Center 902: Molecular principles of RNA-based regulation; R.S. is a recipient of a stipend of the Fonds der Chemischen Industrie; E.C. was supported by a stipend of the Polytechnische Gesellschaft; H.R.A. Jonker is supported by DFG in Forschergruppe FOR2509; H.S. and B.F. are supported by the DFG in graduate school CLIC [GRK 1986]; Work at BMRZ was supported by the state of Hesse. Funding for open access charge: Goethe University Frankfurt.
VL  - 49.2021
IS  - 13
SP  - 7753
EP  - 7764
PB  - Oxford Univ. Press
CY  - Oxford
ER  -