Assembly and entry mechanisms of Semliki Forest virus.
The alphavirus Semliki Forest (SFV) is an enveloped virus with a positive single-stranded RNA genome. The genome is complexed with 240 copies of a capsid protein into a nucleocapsid structure. In the membrane the virus carries an equal number of copies of a membrane protein heterodimer. The latter oligomers are grouped into clusters of three. These structures form the spikes of the virus and carry its entry functions, that is receptor binding and membrane fusion activity. The membrane protein heterodimer is synthesized as a p62E1 precursor protein which upon transport to the cell surface is cleaved into the mature E2E1 form. Recent studies have given much new information on the assembly and entry mechanism of this simple RNA virus. Much of this work has been possible through the construction of a complete cDNA clone of the SFV genome which can be used for in vitro transcription of infectious RNA. One important finding has been to show that a spike deletion variant and a capsid protein deletion variant are budding-negative when expressed separately but can easily complement each other when transfected into the same cell. This shows clearly that enveloped viruses use different budding strategies: one which depends on a nucleocapsid-spike interaction as exemplified by SFV and another one which is based on a direct core-lipid bilayer interaction as shown before to be the case with retroviruses. Another important finding concerns the activation process of the presumed fusion protein of SFV, the E1 subunit. In the original p62E1 heterodimer E1 is completely inactive. Activation proceeds in several steps. First p62 cleavage activates the potential for low pH inducible fusion. Next the low pH which surrounds incoming virus in endosomes induces dissociation of the heterodimeric structure. This is followed by a rearrangement of E1 subunits into homotrimers which are fusion active.
Department of Molecular Biology, Huddinge, Sweden.
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Garoff
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Wilschut
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Liljeström
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Wahlberg
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Bron
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Suomalainen
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Smyth
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Salminen
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Barth
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Zhao
D000818:Animals; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D006224:Cricetinae; D016679:Genome, Viral; D012367:RNA, Viral / Q000235:genetics; D012672:Semliki forest virus / Q000254:growth & development*; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics; D014760:Viral Fusion Proteins / Q000378:metabolism; D014779:Virus Replication
D012367:RNA, Viral; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins