Characterization of mutant tobacco mosaic virus coat protein that interferes with virus cell-to-cell movement.
Expression of tobacco mosaic virus (TMV) coat protein (CP) in plants confers resistance to infection by TMV and related tobamoviruses. Certain mutants of the CP (CP(T42W)) provide much greater levels of resistance than wild-type (wt) CP. In the present work, infection induced by RNA transcripts of TMV clones that contain wt CP or mutant CP(T42W) fused to the green fluorescent protein (GFP) (TMV-CP:GFP, TMV-CP(T42W):GFP) and clones harboring TMV movement protein (MP):GFP were followed in nontransgenic and transgenic tobacco BY-2 protoplasts and Nicotiana tabaccum Xanthi-nn plants that express wt CP or CP(T42W). On nontransgenic and wt CP transgenic plants, TMV-CP:GFP produced expanding, highly fluorescent disk-shaped areas. On plants expressing CP(T42W), infection by TMV-CP:GFP or TMV-MP:GFP-CP produced infection sites of smaller size that were characterized by low fluorescence, reflecting reduced levels of virus spread and reduced accumulation of both CP:GFP and MP:GFP. TMV-CP(T42W):GFP failed to produce visible infection sites on nontransgenic plants, yet produced normal infection sites on MP-transgenic plants that produce MP. TMV infection of transgenic BY-CP(T42W) protoplasts resulted in very low levels of MP accumulation, whereas on BY-CP protoplasts (containing wt CP), infection produced higher levels of MP than in nontransgenic BY-2 cells. The results suggest that wt CP has a positive effect on the production of MP, whereas the CP(T42W) has a negative effect on MP accumulation and/or function. This effect results in very high levels of resistance to TMV infection in plants containing CP(T42W). This report shows that the CP of a plant virus regulates production of the MP, and that a mutant CP interferes with MP accumulation and cell-to-cell movement of infection.
Bendahmane
Mohammed
M
Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, MO 63132, USA.
Szecsi
Judit
J
Chen
Iju
I
Berg
R Howard
RH
Beachy
Roger N
RN
D019943:Amino Acid Substitution / Q000235:genetics; D002213:Capsid / Q000378:metabolism*; D002460:Cell Line; D009068:Movement; D009154:Mutation / Q000235:genetics*; D030821:Plants, Genetically Modified; D021381:Protein Transport; D020871:RNA Stability; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014026:Nicotiana / Q000821:virology*; D014027:Tobacco Mosaic Virus / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication
D012367:RNA, Viral; D011993:Recombinant Fusion Proteins