title: Studies of coat protein-mediated resistance to tobacco mosaic tobamovirus: correlation between assembly of mutant coat proteins and resistance.
abstract: Coat protein-mediated resistance (CP-MR) has been widely used to protect transgenic plants against virus diseases. To characterize the mechanisms of CP-MR to tobacco mosaic tobamovirus (TMV) we developed mutants of the coat protein that affected subunit-subunit interactions. Mutant CPs were expressed during TMV replication as well as in transgenic Nicotiana tabacum plants. The mutation T42-->W increased protein aggregation and T28-->W abolished aggregation and assembly, while the mutations T28-->W plus T42-->W and T89-->W altered normal CP subunit-subunit interactions. The mutant T28W was unable to assemble virus-like particles (VLPs) during infection and in transgenic plants failed to aggregate; this protein conferred no protection against challenge of transgenic plants by TMV. The mutant T42W had strong CP subunit-subunit interactions and formed VLPs but not infectious virions. Transgenic lines with this protein exhibited stronger protection against TMV infection than transgenic plants that contained the wild-type (wt) CP. It is proposed that increased resistance conferred by the T42W mutant results from strong interaction between transgenic CP subunits and challenge virus CP subunits. CP carrying the mutation T89-->W formed flexuous and unstable VLPs whereas the double mutant T28W:T42W formed open helical structures that accumulated as paracrystalline arrays. In transgenic plants, T89W and the double mutant CPs showed reduced ability to aggregate and provided lower protection against TMV infection than wt CP. A strong correlation between normal CP subunit-subunit interactions and CP-MR is observed, and a model for CP-MR involving interactions between the transgenic CP and the CP of the challenge virus as well as interference with virus movement is discussed.
authors.affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: M
authors.initials: M
authors.lastname: Bendahmane
authors.forename: J H
authors.initials: JH
authors.lastname: Fitchen
authors.forename: G
authors.initials: G
authors.lastname: Zhang
authors.forename: R N
authors.initials: RN
authors.lastname: Beachy
mesh_terms: D002213:Capsid / Q000096:biosynthesis* / Q000737:chemistry / Q000648:ultrastructure; D003196:Computer Graphics; D007113:Immunity, Innate; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D018515:Plant Leaves; D030821:Plants, Genetically Modified / Q000821:virology; D010947:Plants, Toxic; D017354:Point Mutation; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000648:ultrastructure; D014026:Nicotiana / Q000235:genetics / Q000821:virology*; D017898:Tobamovirus / Q000302:isolation & purification / Q000472:pathogenicity* / Q000502:physiology*; D014364:Tryptophan; D014779:Virus Replication
chemical_list: D046911:Macromolecular Substances; D011994:Recombinant Proteins; D014364:Tryptophan