Post-translational modifications of Abutilon mosaic virus movement protein (BC1) in fission yeast.
The movement protein (MP) of Abutilon mosaic virus (AbMV, Geminiviridae) exhibited a complex band pattern upon gel electrophoresis indicating its post-translational modification when expressed in AbMV-infected plants or, ectopically, in fission yeasts. High-resolution separation according to charge and molecular weight in acetic acid/urea polyacrylamide or sodium dodecyl sulphate polyacrylamide gels followed by western blot analysis revealed a pattern of AbMV MP from infected plants more related to that from fission yeast than from bacteria. For this reason, expression in fission yeast was established as an experimental system to study post-translational modifications of AbMV MP. Metabolic labelling with 32P-orthophosphate confirmed a phosphorylation of all MP variants suggesting multiple phosphorylation sites. Treatment with calf intestinal alkaline phosphatase removed this label completely, but was unable to eliminate all protein bands with lower electrophoretic mobility. Thus, multiple phosphorylations contribute to the complex migration behaviour of MP, but additional post-translational modifications may occur as well.
Kleinow
Tatjana
T
Institute of Biology, Department of Molecular Biology and Plant Virology, University of Stuttgart, Pfaffenwaldring 57, 70550 Stuttgart, Germany. tatjana.kleinow@bio.uni-stuttgart.de
Holeiter
Gerlinde
G
Nischang
Marc
M
Stein
Monika
M
Karayavuz
Miriam
M
Wege
Christina
C
Jeske
Holger
H
D004591:Electrophoresis, Polyacrylamide Gel; D017791:Geminiviridae / Q000235:genetics / Q000378:metabolism*; D053830:Plant Viral Movement Proteins / Q000378:metabolism* / Q000502:physiology; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D053830:Plant Viral Movement Proteins; D014764:Viral Proteins