Evidence for proteolytic processing of tobacco mosaic virus movement protein in Arabidopsis thaliana.
Two ecotypes of Arabidopsis thaliana were transformed with the gene encoding tobacco mosaic virus (TMV) movement protein (P30). P30 accumulated largely in a subcellular fraction containing cell wall components and as a soluble protein. The protein migrated in denaturing gels with an M(r) of 30K, significantly faster than P30 (M(r) approximately 34K) accumulating after expression in transgenic tobacco, Escherichia coli or Spodoptera frugiperda cells, or after virus multiplication in tobacco. The P30 from A. thaliana infected with TMV for 14 days comigrated with that from E. coli, but that from A. thaliana infected for 49 days was of the smaller size. The use of antisera specific for the N- or C-termini of P30 showed that in A. thaliana P30 was proteolytically processed at the N-terminus, a region essential for P30 function. The failure of these plants to complement a TMV P30 mutant indicated that processed P30 was nonfunctional, although the processing was not so rapid that it prevented the development of systemic infections with wild type TMV. The absence of detectable P30 phosphorylation in A. thaliana demonstrated that phosphorylation was not essential for movement protein function and suggested that this species may use proteolytic cleavage of the N-terminus as an alternative strategy to tobacco for deactivating P30.
Hughes
R K
RK
Department of Virus Research, John Innes Centre, Colney, Norwich, U.K.
Perbal
M C
MC
Maule
A J
AJ
Hull
R
R
D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010766:Phosphorylation; D053830:Plant Viral Movement Proteins; D030821:Plants, Genetically Modified; D011499:Protein Processing, Post-Translational; D018411:Spodoptera / Q000235:genetics; D013347:Subcellular Fractions / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D053830:Plant Viral Movement Proteins; D014764:Viral Proteins