Self-assembly of double-stranded RNA bacteriophages.
Double-stranded RNA viruses infecting bacterial hosts belong to the Cystoviridae family. Bacteriophage phi6 is one of the best characterized dsRNA viruses and shares structural as well as functional similarities with other well-studied eukaryotic dsRNA viruses (e.g. L-A, rotavirus, bluetongue virus, and reovirus). The assembly pathway of the enveloped, triple-layered phi6 virion has been well documented and can be divided into four distinct steps which are (1) procapsid formation, (2) genome encapsidation and replication, (3) nucleocapsid surface shell assembly, and (4) envelope formation. In this review, we focus primarily on the procapsid and nucleocapsid assembly for which in vitro systems have been established. The in vitro assembly systems have been instrumental in revealing assembly intermediates and conformational changes that are common to phi6 and phi8, two cystoviruses with negligible sequence homology. Two viral enzymes, the packaging NTPase (P4) and the RNA-dependent RNA polymerase (P2), were found essential for the nucleation step. The nucleation complex contains one or more tetramers of the major procapsid protein (P1) and is further stabilized by protein P4. Interaction of P1 and P4 during assembly is accompanied by an additional folding of their respective polypeptide chains. The in vitro assembled procapsids were shown to selectively package and replicate the genomic ssRNA. Furthermore, in vitro assembly of infectious nucleocapsids has been achieved in the case of phi6. The in vitro studies indicate that the nucleocapsid coat protein (P8) assembles around the polymerase complex in a template-assisted manner. Implications for the assembly of other dsRNA viruses are also presented.
Poranen
Minna M
MM
Department of Biological and Environmental Sciences, University of Helsinki, Viikinkaari 5, PL 56, FIN-00014, Finland. minna.poranen@helsinki.fi
Tuma
Roman
R
D017126:Bacteriophage phi 6 / Q000235:genetics / Q000502:physiology; D002213:Capsid / Q000502:physiology; D017910:Cystoviridae / Q000235:genetics / Q000502:physiology*; D008954:Models, Biological; D011995:Recombination, Genetic; D012087:Reoviridae / Q000235:genetics / Q000502:physiology; D019065:Virus Assembly