Early steps in reovirus infection are associated with dramatic changes in supramolecular structure and protein conformation: analysis of virions and subviral particles by cryoelectron microscopy and image reconstruction.
Three structural forms of type 1 Lang reovirus (virions, intermediate subviral particles [ISVPs], and cores) have been examined by cryoelectron microscopy (cryoEM) and image reconstruction at 27 to 32-A resolution. Analysis of the three-dimensional maps and known biochemical composition allows determination of capsid protein location, globular shape, stoichiometry, quaternary organization, and interactions with adjacent capsid proteins. Comparisons of the virion, ISVP and core structures and examination of difference maps reveal dramatic changes in supra-molecular structure and protein conformation that are related to the early steps of reovirus infection. The intact virion (approximately 850-A diam) is designed for environmental stability in which the dsRNA genome is protected not only by tight sigma 3-mu 1, lambda 2-sigma 3, and lambda 2-mu 1 interactions in the outer capsid but also by a densely packed core shell formed primarily by lambda 1 and sigma 2. The segmented genome appears to be packed in a liquid crystalline fashion at radii < 240 A. Depending on viral growth conditions, virions undergo cleavage by enteric or endosomal/lysosomal proteases, to generate the activated ISVP (approximately 800-A diam). This transition involves the release of an outer capsid layer spanning radii from 360 to 427 A that is formed by 60 tetrameric and 60 hexameric clusters of ellipsoidal subunits of sigma 3. The vertex-associated cell attachment protein, sigma 1, also undergoes a striking change from a poorly visualized, more compact form, to an extended, flexible fiber. This conformational change may maximize interactions of sigma 1 with cell surface receptors. Transcription of viral mRNAs is mediated by the core particle (approximately 600-A diam), generated from the ISVP after penetration and uncoating. The transition from ISVP to core involves release of the 12 sigma 1 fibers and the remaining outer capsid layer formed by 200 trimers of rod-shaped mu 1 subunits that span radii from 306 to 395 A. In the virion and ISVP, flower-shaped pentamers of the lambda 2 protein are centered at the vertices. In the ISVP-to-core transition, domains of the lambda 2 subunits rotate and swing upward and outward to form a turret-like structure extending from radii 305 to 400 A, with a diameter of 184 A, and a central channel 84 A wide. This novel conformational change allows the potential diffusion of substrates for transcription and exit of newly synthesized mRNA segments.(ABSTRACT TRUNCATED AT 400 WORDS)
Dryden
K A
KA
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907.
Wang
G
G
Yeager
M
M
Nibert
M L
ML
Coombs
K M
KM
Furlong
D B
DB
Fields
B N
BN
Baker
T S
TS
D000818:Animals; D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*; D002478:Cells, Cultured; D003080:Cold Temperature; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000382:microbiology; D007091:Image Processing, Computer-Assisted; D046911:Macromolecular Substances; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods; D011487:Protein Conformation; D012330:RNA, Double-Stranded / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012087:Reoviridae / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure*; D012088:Reoviridae Infections / Q000378:metabolism / Q000503:physiopathology*; D014158:Transcription, Genetic; D014758:Viral Core Proteins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*; D014771:Virion / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*
D004279:DNA, Viral; D046911:Macromolecular Substances; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D012367:RNA, Viral; D014758:Viral Core Proteins