Analyses of the effects of all ubiquitin point mutants on yeast growth rate.
Analyses of the effects of all ubiquitin point mutants on yeast growth rate.
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DOI：
10.1016/j.jmb.2013.01.032
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发表时间：
2013-04-26
影响因子：
5.6
通讯作者：
Bolon, Daniel N. A.
中科院分区：
文献类型：
--
作者：
Roscoe, Benjamin P.;Thayer, Kelly M.;Zeldovich, Konstantin B.;Fushman, David;Bolon, Daniel N. A.
蛋白质的氨基酸序列决定其功能。我们使用批量竞争和聚焦深度测序来研究所有泛素点突变体对酵母生长速率的影响。泛素功能的许多方面已经被仔细研究，这使得我们的生长分析的解释，在丰富的结构，生物物理和生物化学知识基础。在泛素表面的一个高度敏感的簇中，几乎每一个氨基酸取代都会导致生长缺陷。相反，相反的面孔几乎可以容忍所有可能的替换。这两个面之间的表面位置表现出中间的突变耐受性。敏感面对应于许多结合伴侣的已知界面。在所有的表面位置，我们观察到埋葬在结构特征的接口和氨基酸取代的数量与强劲的增长兼容之间的强相关性。这一结果表明，结合是泛素功能的主要决定因素。在溶剂不可及的核心泛素的所有位置容忍有限数量的取代，与疏水性氨基酸特别是可互换的。一些突变无效的酵母生长先前显示填充折叠构象，这表明对于这些突变体，构象的细微变化导致功能缺陷。核心内对突变最敏感的区域位于C-末端附近，这是许多关键结合配偶体的焦点结合位点。这些结果表明，核心突变可能经常通过对结构或动力学的细微干扰导致功能缺陷。
The amino acid sequence of a protein governs its function. We used bulk competition and focused deep sequencing to investigate the effects of all ubiquitin point mutants on yeast growth rate. Many aspects of ubiquitin function have been carefully studied, which enabled interpretation of our growth analyses in light of a rich structural, biophysical and biochemical knowledge base. In one highly sensitive cluster on the surface of ubiquitin almost every amino acid substitution caused growth defects. In contrast, the opposite face tolerated virtually all possible substitutions. Surface locations between these two faces exhibited intermediate mutational tolerance. The sensitive face corresponds to the known interface for many binding partners. Across all surface positions, we observe a strong correlation between burial at structurally characterized interfaces and the number of amino acid substitutions compatible with robust growth. This result indicates that binding is a dominant determinant of ubiquitin function. In the solvent inaccessible core of ubiquitin all positions tolerated a limited number of substitutions, with hydrophobic amino acids especially interchangeable. Some mutations null for yeast growth were previously shown to populate folded conformations indicating that for these mutants subtle changes to conformation caused functional defects. The most sensitive region to mutation within the core was located near the C-terminus that is a focal binding site for many critical binding partners. These results indicate that core mutations may frequently cause functional defects through subtle disturbances to structure or dynamics.
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影响因子：
56.9
作者：
Dahiyat, BI;Mayo, SL
通讯作者：
Mayo, SL
影响因子：
56.9
作者：
CONNOLLY, ML
通讯作者：
CONNOLLY, ML
影响因子：
2.9
作者：
Benítez-Cardoza, CG;Stott, K;Jackson, SE
通讯作者：
Jackson, SE
DOI：
10.1073/pnas.1209751109
发表时间：
2012-10-16
影响因子：
11.1
作者：
Araya, Carlos L.;Fowler, Douglas M.;Fields, Stanley
通讯作者：
Fields, Stanley
影响因子：
4.5
作者：
Domingo-Calap P;Cuevas JM;Sanjuán R
通讯作者：
Sanjuán R