Structure of a pentameric virion-associated fiber with a potential role in Orsay virus entry to host cells

PLoS Pathogens

Yusong R. Guo¹  Yizhi J. Tao¹  Wang Yuan¹  Yanlin Fan¹  Ying Zhou¹  Weiwei Zhong²  Borries Demeler³  Matthew V. Holt⁴  Tao Wang⁴  Nicolas L. Young⁴ 

[1] Department of BioSciences, Rice University, MS-140, Houston, Texas, United States of America;Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, United States of America;The University of Texas Health Science Center at San Antonio, Department of Biochemistry, MC 7760, 7703 Floyd Curl Drive, San Antonio, Texas, United States of America;Verna & Marrs McLean Department of Biochemistry & Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, United States of America

关键词: Viral packaging;    Crystal structure;    Caenorhabditis elegans;    Nematode infections;    Viral structure;    RNA interference;    Recombinant proteins;    Protein structure;   

DOI  :  10.1371/journal.ppat.1006231

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Despite the wide use of Caenorhabditis elegans as a model organism, the first virus naturally infecting this organism was not discovered until six years ago. The Orsay virus and its related nematode viruses have a positive-sense RNA genome, encoding three proteins: CP, RdRP, and a novel δ protein that shares no homology with any other proteins. δ can be expressed either as a free δ or a CP-δ fusion protein by ribosomal frameshift, but the structure and function of both δ and CP-δ remain unknown. Using a combination of electron microscopy, X-ray crystallography, computational and biophysical analyses, here we show that the Orsay δ protein forms a ~420-Å long, pentameric fiber with an N-terminal α-helical bundle, a β-stranded filament in the middle, and a C-terminal head domain. The pentameric nature of the δ fiber has been independently confirmed by both mass spectrometry and analytical ultracentrifugation. Recombinant Orsay capsid containing CP-δ shows protruding long fibers with globular heads at the distal end. Mutant viruses with disrupted CP-δ fibers were generated by organism-based reverse genetics. These viruses were found to be either non-viable or with poor infectivity according to phenotypic and qRT-PCR analyses. Furthermore, addition of purified δ proteins to worm culture greatly reduced Orsay infectivity in a sequence-specific manner. Based on the structure resemblance between the Orsay CP-δ fiber and the fibers from reovirus and adenovirus, we propose that CP-δ functions as a cell attachment protein to mediate Orsay entry into worm intestine cells.

【 授权许可】

CC BY