In vitro properties of hordeivirus TGB1 protein forming ribonucleoprotein complexes.
Hordeivirus movement protein encoded by the first gene of the triple gene block (TGB1 protein, TGBp1) interacts in vivo with viral genomic and subgenomic RNAs to form ribonucleoprotein (RNP) particles that are considered to be a form of viral genome (non-virion transport form) capable of cell-to-cell and long-distance transport in infected plants. The structures of these RNPs have not been elucidated. The poa semilatent virus (PSLV) TGBp1 contains a structured C-terminal NTPase/helicase domain and an N-terminal extension region consisting of two domains - a completely intrinsically disordered extreme N-terminal domain and an internal domain (ID) with structure resembling a partially disordered molten globule. Here, we characterized the structures assembled in vitro by the full-length PSLV TGBp1 alone or in the presence of viral RNA. The PSLV TGBp1 was capable of multimerization and self-assembly into extended high-molecular-mass complexes. These complexes disassembled to apparent monomers upon incubation with ATP. Upon incubation with viral RNA, the PSLV TGBp1 in vitro formed RNP structures that appeared as filamentous particles resembling virions of helical filamentous plant viruses in morphology and dimensions. By comparing the biophysical characteristics of PSLV TGBp1 and its domains in the presence and absence of RNA, we show that the ID plays the main structural role in the self-interactions and RNA interactions of TGBp1 leading to the assembly of virus-like RNP particles.
A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Leninsky Gory, Moscow 119992, Russia.
Valentin V
VV
Makarov
Department of Biology, Lomonosov Moscow State University, Leninsky Gory, Moscow 119992, Russia.
Svetlana S
SS
Makarova
A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Leninsky Gory, Moscow 119992, Russia.
Antonida V
AV
Makhotenko
M. M. Shemyakin and Yu. A. Ovchinnikov Bioorganic Chemistry Institute, Miklukho-Maklaya str. 16/10, Moscow 117997, Russia.
Ekaterina A
EA
Obraztsova
A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Leninsky Gory, Moscow 119992, Russia.
Natalia O
NO
Kalinina
D016679:Genome, Viral; D012328:RNA Viruses / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins