Homotypic interaction and multimerization of nucleocapsid protein of tomato spotted wilt tospovirus: identification and characterization of two interacting domains.
The nucleocapsid protein (N) of tomato spotted wilt tospovirus (TSWV) plays a central role in the viral life cycle. With the aid of the yeast two-hybrid system and surface plasmon resonance analysis, homotypic interaction and multimerization of the N protein was detected. Analysis of deletion mutants identified two binding regions in the protein, located at the N terminus (amino acids 1-39) and the C terminus (amino acids 233-248), respectively, implying a "head-to-tail" interaction of the N terminus with the C terminus to form a multimeric chain. Further characterization of the binding domains was performed by site-directed mutagenesis. Two phenylalanines (F242 and F246) highly conserved in the N proteins within the Tospovirus genus were shown to play a crucial role in the interaction.
Uhrig
J F
JF
Max-Planck-Institut für Züchtungsforschung, Abteilung Genetische Grundlagen der Pflanzenzüchtung, Carl-von-Linné-Weg 10, D-50829 Cologne, Germany.
Soellick
T R
TR
Minke
C J
CJ
Philipp
C
C
Kellmann
J W
JW
Schreier
P H
PH
D000595:Amino Acid Sequence; D001665:Binding Sites; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance; D017865:Tospovirus
D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins; C099603:nucleocapsid protein, Tospovirus