Katz Garrett, Benkarroum Younes, Wei Hui, Rice William J, Bucher Doris, Alimova Alexandra, Katz Al, Klukowska Joanna, Herman Gabor T, Gottlieb Paul
Department of Microbiology and Immunology, Sophie Davis School of Biomedical Education, The City College of New York, New York, New York, United States of America.
Department of Computer Science, The Graduate Center, City University of New York, New York, New York, United States of America.
PLoS One. 2014 Feb 6;9(2):e88288. doi: 10.1371/journal.pone.0088288. eCollection 2014.
Cryo-electron microscopy projection image analysis and tomography is used to describe the overall architecture of influenza B/Lee/40. Algebraic reconstruction techniques with utilization of volume elements (blobs) are employed to reconstruct tomograms of this pleomorphic virus and distinguish viral surface spikes. The purpose of this research is to examine the architecture of influenza type B virions by cryo-electron tomography and projection image analysis. The aims are to explore the degree of ribonucleoprotein disorder in irregular shaped virions; and to quantify the number and distribution of glycoprotein surface spikes (hemagglutinin and neuraminidase) on influenza B. Projection image analysis of virion morphology shows that the majority (∼83%) of virions are spherical with an average diameter of 134±19 nm. The aspherical virions are larger (average diameter = 155±47 nm), exhibit disruption of the ribonucleoproteins, and show a partial loss of surface protein spikes. A count of glycoprotein spikes indicates that a typical 130 nm diameter type B virion contains ∼460 surface spikes. Configuration of the ribonucleoproteins and surface glycoprotein spikes are visualized in tomogram reconstructions and EM densities visualize extensions of the spikes into the matrix. The importance of the viral matrix in organization of virus structure through interaction with the ribonucleoproteins and the anchoring of the glycoprotein spikes to the matrix is demonstrated.
冷冻电子显微镜投影图像分析和断层扫描用于描述乙型流感病毒/李/40株的整体结构。利用体素(团块)的代数重建技术用于重建这种多形病毒的断层图像,并区分病毒表面的刺突。本研究的目的是通过冷冻电子断层扫描和投影图像分析来研究乙型流感病毒粒子的结构。其目标是探究不规则形状病毒粒子中核糖核蛋白的无序程度;并量化乙型流感病毒糖蛋白表面刺突(血凝素和神经氨酸酶)的数量和分布。病毒粒子形态的投影图像分析表明,大多数(约83%)病毒粒子呈球形,平均直径为134±19纳米。非球形病毒粒子更大(平均直径 = 155±47纳米),表现出核糖核蛋白的破坏,并显示表面蛋白刺突部分缺失。糖蛋白刺突的计数表明,典型的直径为130纳米的乙型病毒粒子含有约460个表面刺突。在断层图像重建中可以看到核糖核蛋白和表面糖蛋白刺突的形态,并且电子显微镜密度显示刺突延伸到基质中。研究证明了病毒基质在通过与核糖核蛋白相互作用来组织病毒结构以及将糖蛋白刺突锚定到基质中的重要性。