美国加州大学旧金山分校Nevan J. Krogan等研究人员合作发现，遗传相互作用图谱可确定蛋白质复合物的整合结构。这一研究成果于2020年12月11日发表在国际学术期刊《科学》上。
研究人员表示，确定蛋白质复合物的结构对于理解细胞功能至关重要。
研究人员描述了一种依赖于体内遗传相互作用测量的整体结构确定方法。研究人员针对与基因缺失交叉或暴露于环境扰动的点突变来构建了表型图谱，然后将两个图谱之间的相似性转换为突变残基之间距离的上限。基于350个突变体的约500,000遗传相互作用，研究人员确定酵母组蛋白H3-H4复合物的结构。
然后，研究人员将该方法应用于酵母RNA聚合酶II的亚基Rpb1-Rpb2和细菌RNA聚合酶的RpoB-RpoC亚基。其精度可与基于化学交联的精度相媲美；同时使用遗传相互作用和交联的限制条件可以进一步提高模型的准确性和精确度。该方法提供了一种有效的手段，可以通过体内观察来增强整体结构的确定。
附：英文原文
Title: Genetic interaction mapping informs integrative structure determination of protein complexes
Author: Hannes Braberg, Ignacia Echeverria, Stefan Bohn, Peter Cimermancic, Anthony Shiver, Richard Alexander, Jiewei Xu, Michael Shales, Raghuvar Dronamraju, Shuangying Jiang, Gajendradhar Dwivedi, Derek Bogdanoff, Kaitlin K. Chaung, Ruth Hüttenhain, Shuyi Wang, David Mavor, Riccardo Pellarin, Dina Schneidman, Joel S. Bader, James S. Fraser, John Morris, James E. Haber, Brian D. Strahl, Carol A. Gross, Junbiao Dai, Jef D. Boeke, Andrej Sali, Nevan J. Krogan
Issue&Volume: 2020/12/11
Abstract: Determining structures of protein complexes is crucial for understanding cellular functions. Here, we describe an integrative structure determination approach that relies on in vivo measurements of genetic interactions. We construct phenotypic profiles for point mutations crossed against gene deletions or exposed to environmental perturbations, followed by converting similarities between two profiles into an upper bound on the distance between the mutated residues. We determine the structure of the yeast histone H3-H4 complex based on ~500,000 genetic interactions of 350 mutants. We then apply the method to subunits Rpb1-Rpb2 of yeast RNA polymerase II and subunits RpoB-RpoC of bacterial RNA polymerase. The accuracy is comparable to that based on chemical cross-links; using restraints from both genetic interactions and cross-links further improves model accuracy and precision. The approach provides an efficient means to augment integrative structure determination with in vivo observations.
DOI: 10.1126/science.aaz4910
Source: https://science.sciencemag.org/content/370/6522/eaaz4910