Department of Biochemistry, University of Oxford, Oxford, United Kingdom; Faculty of Science, Universiti Brunei Darussalam, Gadong, Brunei Darussalam.
Department of Biochemistry, University of Oxford, Oxford, United Kingdom.
J Biol Chem. 2021 Jan-Jun;296:100316. doi: 10.1016/j.jbc.2021.100316. Epub 2021 Jan 29.
A central role for the influenza matrix protein 1 (M1) is to form a polymeric coat on the inner leaflet of the host membrane that ultimately provides shape and stability to the virion. M1 polymerizes upon binding membranes, but triggers for conversion of M1 from a water-soluble component of the nucleus and cytosol into an oligomer at the membrane surface are unknown. While full-length M1 is required for virus viability, the N-terminal domain (M1NT) retains membrane binding and pH-dependent oligomerization. We studied the structural plasticity and oligomerization of M1NT in solution using NMR spectroscopy. We show that the isolated domain can be induced by sterol-containing compounds to undergo a conformational change and self-associate in a pH-dependent manner consistent with the stacked dimer oligomeric interface. Surface-exposed residues at one of the stacked dimer interfaces are most sensitive to sterols. Several perturbed residues are at the interface between the N-terminal subdomains and are also perturbed by changes in pH. The effects of sterols appear to be indirect and most likely mediated by reduction in water activity. The local changes are centered on strictly conserved residues and consistent with a priming of the N-terminal domain for polymerization. We hypothesize that M1NT is sensitive to changes in the aqueous environment and that this sensitivity is part of a mechanism for restricting polymerization to the membrane surface. Structural models combined with information from chemical shift perturbations indicate mechanisms by which conformational changes can be transmitted from one polymerization interface to the other.
流感基质蛋白 1(M1)的一个核心作用是在内膜层形成聚合外壳,最终为病毒提供形状和稳定性。M1 在结合膜时聚合,但将 M1 从核和细胞质中的水溶性成分转换为膜表面低聚物的触发因素尚不清楚。虽然全长 M1 是病毒存活所必需的,但 N 端结构域(M1NT)保留了膜结合和 pH 依赖性寡聚化的能力。我们使用 NMR 光谱研究了 M1NT 在溶液中的结构可塑性和寡聚化。我们表明,分离的结构域可以被含有固醇的化合物诱导,发生构象变化,并以 pH 依赖性的方式自组装,与堆叠二聚体寡聚界面一致。堆叠二聚体界面之一上的表面暴露残基对固醇最敏感。几个扰动的残基位于 N 端亚结构域之间的界面上,并且 pH 变化也会受到影响。固醇的影响似乎是间接的,很可能是通过降低水活度来介导的。局部变化集中在严格保守的残基上,与 N 端结构域聚合的启动一致。我们假设 M1NT 对水相环境的变化敏感,这种敏感性是限制聚合在膜表面的机制的一部分。结构模型结合化学位移扰动信息表明了构象变化可以从一个聚合界面传递到另一个聚合界面的机制。