caa a22 4500 445325232 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9573-6 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9573-6 Interactions of protein side chains with RNA defined with REDOR solid state NMR [Elektronische Daten] [Wei Huang, Gabriele Varani, Gary Drobny] Formation of the complex between human immunodeficiency virus type-1 Tat protein and the transactivation response region (TAR) RNA is vital for transcriptional elongation, yet the structure of the Tat-TAR complex remains to be established. The NMR structures of free TAR, and TAR bound to Tat-derived peptides have been obtained by solution NMR, but only a small number of intermolecular NOEs could be identified unambiguously, preventing the determination of a complete structure. Here we show that a combination of multiple solid state NMR REDOR experiments can be used to obtain multiple distance constraints from 15N to 13C spins within the backbone and side chain guanidinium groups of arginine in a Tat-derived peptide, using 19F spins incorporated into the base of U23 in TAR and 31P spins in the P22 and P23 phosphate groups. Distances between the side chain of Arg52 and the base and phosphodiester backbone near U23 measured by REDOR NMR are comparable to distances observed in solution NMR-derived structural models, indicating that interactions of TAR RNA with key amino acid side chains in Tat are the same in the amorphous solid state as in solution. This method is generally applicable to other protein-RNA complexes where crystallization or solution NMR has failed to provide high resolution structural information. Springer Science+Business Media B.V., 2011 Solid state NMR nationallicence REDOR nationallicence RNA nationallicence Tat-TAR nationallicence Huang Wei Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut Varani Gabriele Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut Drobny Gary Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 347-356 0925-2738 51:3<347 2011 51 10858 https://doi.org/10.1007/s10858-011-9573-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9573-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Huang Wei Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut NATIONALLICENCE 700 1- Varani Gabriele Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut NATIONALLICENCE 700 1- Drobny Gary Department of Chemistry, University of Washington, Box 351700, 98195, Seattle, Washington, DC, USA aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 347-356 0925-2738 51:3<347 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer