[Use of three-hybrid system to detect RNA-binding activity of alfalfa mosaic virus coat protein].
We used yeast three-hybrid system, for studying interaction of alfalfa mosaic virus coat protein AMVCP (AMVCP) with RNA4, which codes this protein. We have shown that AMVCP with high affinity is bound to plus-chain of RNA4 in vivo. The mutational analysis has shown, that the N-terminal part of AMVCP (aa 1 to 85) contains RNA-binding domain. C-terminal part of this protein (aa 86 to 221) does not participate in direct interaction with RNA4. However activity of the reporter-gene LacZ, which codes beta-galactosidase, in case of interaction only N-terminal part of AMVCP is five times lower, in comparison with full-length hybrid protein, that confirms that the tertiary structure of full-length AMVCP is more favourable for interaction with RNA4.
Spiridonov
V G
VG
National Agricultural University, Kyiv, Ukraine.
Smirnova
S A
SA
Mel'nichuk
M D
MD
D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D017930:Genes, Reporter / Q000235:genetics; D007763:Lac Operon / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; C095691:coat protein, Alfalfa mosaic virus; D001616:beta-Galactosidase