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PMID: 11160687 Published · ppublish English Journal Article

Alfalfa mosaic virus replicase proteins P1 and P2 interact and colocalize at the vacuolar membrane.

Journal of virology ·Vol. 75 ·No. 4 ·2001-02-00 ·Pages 1879-87

Van Der Heijden MW, Carette JE, Reinhoud PJ, Haegi A, Bol JF

Abstract

Replication of Alfalfa mosaic virus (AMV) RNAs depends on the virus-encoded proteins P1 and P2. P1 contains methyltransferase- and helicase-like domains, and P2 contains a polymerase-like domain. Coimmunoprecipitation experiments revealed an interaction between in vitro translated-P1 and P2 and showed that these proteins are present together in fractions with RNA-dependent RNA polymerase activity. A deletion analysis in the yeast two-hybrid system showed that in P1 the C-terminal sequence of 509 amino acids with the helicase domain was necessary for the interaction. In P2, the sequence of the N-terminal 241 aa was required for the interaction. In infected protoplasts, P1 and P2 colocalized at a membrane structure that was identified as the tonoplast (i.e., the membrane that surrounds the vacuoles) by using a tonoplast intrinsic protein as a marker in immunofluorescence studies. While P1 was exclusively localized on the tonoplast, P2 was found both at the tonoplast and at other locations in the cell. As Brome mosaic virus replication complexes have been found to be associated with the endoplasmic reticulum (M. A. Restrepo-Hartwig and P. Ahlquist, J. Virol. 70:8908-8916, 1996), viruses in the family Bromoviridae apparently select different cellular membranes for the assembly of their replication complexes.

MeSH Terms
Alfalfa mosaic virus/enzymology,genetics,physiology Intracellular Membranes/enzymology,virology Methyltransferases/genetics,metabolism Plants/virology Precipitin Tests Protoplasts/virology RNA Helicases/genetics,metabolism RNA-Dependent RNA Polymerase/genetics,metabolism Two-Hybrid System Techniques Vacuoles/enzymology,virology Viral Proteins/genetics,metabolism Virus Replication
Chemicals
Viral Proteins Methyltransferases RNA-Dependent RNA Polymerase RNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Van Der Heijden M W
Institute of Molecular Plant Sciences, Gorlaeus Laboratories, Leiden University, 2300 RA Leiden, The Netherlands.
Carette J E
Reinhoud P J
Haegi A
Bol J F
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.75.4.1879-1887.2001
Published
2001-02-00
Pages
1879-87
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC114098
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.75.4.1879-1887.2001 PMC Full Text View journal details
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