Cell-to-cell trafficking, subcellular distribution, and binding to coat protein of Broad bean wilt virus 2 VP37 protein.
Broad bean wilt virus 2 (BBWV 2) is a member of the genus Fabavirus of the family Comoviridae. To date, a movement protein (MP) of BBWV 2 has not been described. Here we demonstrate that the green fluorescent protein (GFP)-VP37 fusion protein can move from initial bombarded cells to neighboring cells in Nicotiana benthamiana epidermal leaves. In addition, the GFP-VP37 fusion protein localizes as a halo around the nucleus and as punctate spots on the cell periphery in N. benthamiana epidermal leaf cells and BY-2 suspension cells. Fluorescence near the nucleus also was co-localized with the endoplasmic reticulum in BY-2 cells. Fibrous networks were found in GFP-VP37 agro-infiltrated N. benthamiana epidermal leaf cells. Deletion analyses indicated that the C-terminal region of the VP37 protein is essential for localization at the cell periphery. Using a blot overlay assay and bimolecular fluorescence complementation assay, the purified 6xHis-tagged VP37 protein was shown to bind specifically to the small coat protein of BBWV 2. The above results indicate that VP37 is a movement protein.
Liu
Chengke
C
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, People's Republic of China.
Meng
Chunmei
C
Xie
Li
L
Hong
Jian
J
Zhou
Xueping
X
D036022:Capsid Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D019181:Fabavirus / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D008856:Microscopy, Fluorescence; D009068:Movement; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000378:metabolism / Q000821:virology; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014026:Nicotiana / Q000378:metabolism / Q000821:virology; D014764:Viral Proteins / Q000378:metabolism*
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins