The TMV movement protein: role of the C-terminal 73 amino acids in subcellular localization and function.
The role of the C-terminal one-third of the tobacco mosaic virus (TMV) 30-kDa movement protein (MP) on its subcellular localization and on virus spread was investigated. We have constructed eight cDNAs encoding MPs with variable size deletions from the C-terminal end. Expression of the truncated proteins was verified in recombinant yeast using an antiserum directed to a synthetic peptide corresponding to 21 amino acids near the N-terminal end of the MP. In transgenic tobacco plants, MP from which more than 55 amino acids were deleted no longer accumulated in the cell wall fraction of a cellular extract, where the complete MP accumulates. Dye diffusion studies showed that both unmodified and modified MPs that accumulate in the cell wall fraction are able to alter plasmodesmatal size exclusion limits. Biological function of the modified MPs was tested in the transgenic plants with the TMV thermosensitive mutant Ls1 and a TMV genomic RNA transcript lacking a functional MP. There was a correlation between the cell wall localization of the modified MPs and its ability to potentiate virus spread. The results presented here demonstrate the dispensability of the C-terminal 55 amino acids of the MP in its subcellular localization in tobacco plants and its role in virus movement. Moreover, our results show that a stretch of 19 amino acids (195 to 213) is essential for localization of the MP to the cell wall fraction of plant cells.
Berna
A
A
Université Laval, Département de Biochimie, Ste-Foy, Québec, Canada.
Gafny
R
R
Wolf
S
S
Lucas
W J
WJ
Holt
C A
CA
Beachy
R N
RN
D002213:Capsid / Q000502:physiology*; D002473:Cell Wall / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D015870:Gene Expression; D005816:Genetic Complementation Test; D010935:Plant Diseases; D053830:Plant Viral Movement Proteins; D010947:Plants, Toxic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013329:Structure-Activity Relationship; D014026:Nicotiana; D014027:Tobacco Mosaic Virus / Q000502:physiology*; D014758:Viral Core Proteins / Q000502:physiology*; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
D053830:Plant Viral Movement Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins; D004247:DNA