Rice black-streaked dwarf virus outer capsid protein P10 has self-interactions and forms oligomeric complexes in solution.
The P10 protein encoded by S10 ORF of Rice black-streaked dwarf virus (RBSDV) was thought to be the component of outer shell of viral particle. In the present study, P10 has an ability for self-interaction as shown by a GAL4 transcription activator-based yeast two-hybrid assay system and further confirmed by in vitro far-Western blot analysis. The domain responsible for P10-P10 self-interaction was mapped to the first 230 amino acids at the N-terminal region of the protein. The oligomerization property of P10 was further investigated using chemical cross-linking with purified recombinant P10 proteins expressed in a baculovirus expression system and glutaraldehyde. Intact P10 recombinants existed predominantly as trimers in solution in the absence of other viral proteins and displayed the oligomeric nature common to all known second-layer protein units of the Reoviridae. A truncated P10 mutant encoding the first 230 N-terminal amino acids lost its ability to form trimers even though dimeric forms were detected during the cross-linking assay. Polyacrylamide gel electrophoresis under reducing or non-reducing conditions suggested that P10 subunits were oligomerized not through intermolecular disulfide bonds, but perhaps through some other type of association, such as hydrophobic or charge interactions.
Liu
Huijun
H
Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology, The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China.
Wei
Chunhong
C
Zhong
Yongwang
Y
Li
Yi
Y
D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D012275:Oryza / Q000821:virology*; D010935:Plant Diseases / Q000821:virology; D020033:Protein Isoforms; D012087:Reoviridae / Q000737:chemistry*
D036022:Capsid Proteins; D020033:Protein Isoforms; D005982:Glutathione Transferase