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GrainGenes Reference Report: JGV-83-443

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Reference	JGV-83-443
Title	Mapping of the P1 proteinase cleavage site in the polyprotein of Wheat streak mosaic virus (genus Tritimovirus)
Journal	Journal of General Virology
Year	2002
Volume	83
Pages	443-450
Author	Choi IR Horken KM Stenger DC French R
Abstract	Summary: Monopartite members of the family Potyviridae utilize three virus-encoded proteinases to cleave the viral polyprotein into mature proteins. The amino-terminal region of the viral polyprotein is autolytically cleaved by the P1 proteinase. A domain required for P1 proteinase activity of Wheat streak mosaic virus (WSMV) was mapped using a series of templates with nested 3'-truncations or 5'-deletions to program in vitro transcription-translation reactions. The WSMV P1 proteinase cleavage site was mapped to a position downstream of amino acid residue 348 and upstream of amino acid residue 353, with the peptide bond between amino acid residues Y352 and G353 the most probable site of hydrolysis. An alignment of potyvirus polyprotein sequences in the carboxy-terminal region of the P1 domain revealed WSMV P1 contained conserved H257, D267, S303 and FIVXG325-329 residues upstream of the cleavage site that are typical of serine proteinases and shown by others to be required for P1 proteolysis in Tobacco etch virus. Insertion of the GUS reporter gene immediately downstream of the P1 cleavage site in a full-length clone of WSMV resulted in systemic infection and GUS expression upon inoculation of plants with in vitro transcripts. When cleaved by P1 at the amino terminus and NIa proteinase at a site engineered in the carboxy-terminus, actsive GUS protein expressed by WSMV in infected wheat had electrophoretic mobility similar to wild-type GUS protein
External Databases	Pubmed: 11807238
Keyword	[ Hide all but 1 of 52 ] acid alignment amino acid amino-terminus cleavage clone complete nucleotide-sequence domain family family potyviridae gene genus gus gus expression hydrolysis in vitro infection inoculation insertion mapping member microbiology mobility mosaic-virus p1 peptide plant potyvirus genome position potyviridae potyvirus program protein proteinase proteolysis region reporter reporter gene residue rymovirus sequence serine serine proteinases site template tobacco tobacco etch virus transcript virus vitro wheat streak mosaic virus wild-type wsmv

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