2M3F

NMR structure of Rsa1p238-259 from S. Cerevisiae

Summary for 2M3F

• Entry DOI: 10.2210/pdb2m3f/pdb
• NMR Information: BMRB: 18959
• Descriptor: Ribosome assembly 1 protein (1 entity in total)
• Functional Keywords: helix, rna binding protein
• Biological source: Saccharomyces cerevisiae (Baker's yeast)
• Cellular location: Nucleus: Q08932
• Total number of polymer chains: 1
• Total formula weight: 2840.32

Authors

Quinternet, M.,Manival, X. (deposition date: 2013-01-17, release date: 2013-11-27, Last modification date: 2024-05-15)

Primary citation

Rothe, B.,Back, R.,Quinternet, M.,Bizarro, J.,Robert, M.C.,Blaud, M.,Romier, C.,Manival, X.,Charpentier, B.,Bertrand, E.,Branlant, C.
Characterization of the interaction between protein Snu13p/15.5K and the Rsa1p/NUFIP factor and demonstration of its functional importance for snoRNP assembly.
Nucleic Acids Res., 42:2015-2036, 2014

PubMed Abstract: The yeast Snu13p protein and its 15.5K human homolog both bind U4 snRNA and box C/D snoRNAs. They also bind the Rsa1p/NUFIP assembly factor, proposed to scaffold immature snoRNPs and to recruit the Hsp90-R2TP chaperone complex. However, the nature of the Snu13p/15.5K-Rsa1p/NUFIP interaction and its exact role in snoRNP assembly remained to be elucidated. By using biophysical, molecular and imaging approaches, here, we identify residues needed for Snu13p/15.5K-Rsa1p/NUFIP interaction. By NMR structure determination and docking approaches, we built a 3D model of the Snup13p-Rsa1p interface, suggesting that residues R249, R246 and K250 in Rsa1p and E72 and D73 in Snu13p form a network of electrostatic interactions shielded from the solvent by hydrophobic residues from both proteins and that residue W253 of Rsa1p is inserted in a hydrophobic cavity of Snu13p. Individual mutations of residues in yeast demonstrate the functional importance of the predicted interactions for both cell growth and snoRNP formation. Using archaeal box C/D sRNP 3D structures as templates, the association of Snu13p with Rsa1p is predicted to be exclusive of interactions in active snoRNPs. Rsa1p and NUFIP may thus prevent premature activity of pre-snoRNPs, and their removal may be a key step for active snoRNP production.

PubMed: 24234454
DOI: 10.1093/nar/gkt1091

Experimental method

SOLUTION NMR