Characterization of the interaction between protein Snu13p/15.5K and the Rsa1p/NUFIP factor and demonstration of its functional importance for snoRNP assembly.
Characterization of the interaction between protein Snu13p/15.5K and the Rsa1p/NUFIP factor and demonstration of its functional importance for snoRNP assembly.
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DOI：
10.1093/nar/gkt1091
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发表时间：
2014-02
影响因子：
14.9
通讯作者：
Branlant C
中科院分区：
文献类型：
--
作者：
Rothé B;Back R;Quinternet M;Bizarro J;Robert MC;Blaud M;Romier C;Manival X;Charpentier B;Bertrand E;Branlant C
酵母Snu13p蛋白及其15.5K的人类同源物都能结合U4 snRNA和box C/D snoRNAs。它们还结合Rsa1p/NUFIP组装因子，被认为可以支撑未成熟的snornp并招募Hsp90-R2TP伴侣复合物。然而，Snu13p/ 15.5K-Rsa1p /NUFIP相互作用的性质及其在snoRNP组装中的确切作用仍有待阐明。通过生物物理、分子和成像方法，我们鉴定了Snu13p/ 15.5K-Rsa1p /NUFIP相互作用所需的残基。通过核磁共振结构测定和对接方法，我们建立了Snup13p-Rsa1p界面的三维模型，表明Rsa1p中的R249、R246和K250残基和Snu13p中的E72和D73残基形成了一个静电相互作用网络，这两个蛋白的疏水残基屏蔽了溶剂，Rsa1p的残基W253插入到Snu13p的疏水腔中。酵母残基的个体突变证明了预测的相互作用对细胞生长和snoRNP形成的功能重要性。使用古细菌盒C/D sRNP三维结构作为模板，预测Snu13p与Rsa1p的关联不排除活性snornp的相互作用。因此，Rsa1p和NUFIP可能会阻止前snoRNP的过早活性，它们的去除可能是活性snoRNP产生的关键步骤。
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.
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DOI：
10.1083/jcb.200601105
发表时间：
2006-04-24
期刊：
The Journal of cell biology
影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
Moras, D
DOI：
10.1083/jcb.200708110
发表时间：
2008-02-11
期刊：
The Journal of cell biology
影响因子：
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通讯作者：
Charpentier B