Interaction of Mal de Río Cuarto virus ( Fijivirus genus) proteins and identification of putative factors determining viroplasm formation and decay

Autor: Gabriela Llauger, Victoria Alfonso, Mariana del Vas, Luis Alejandro de Haro
Rok vydání: 2017
Předmět:
0301 basic medicine
Cancer Research
Agroinfiltration
Otras Ciencias Biológicas
Amino Acid Motifs
Gene Expression
Nicotiana benthamiana
Viral Nonstructural Proteins
Reoviridae
Ciencias Biológicas
Serine
03 medical and health sciences
Bimolecular fluorescence complementation
Two-Hybrid System Techniques
Virology
Protein Interaction Mapping
Tobacco
MRCV
PEST SEQUENCE
Viroplasm
Protein Interaction Domains and Motifs
Amino Acid Sequence
Threonine
Conserved Sequence
Plant Diseases
Inclusion Bodies
Sequence Homology
Amino Acid
Y2H
biology
Protoplasts
Agricultura
Oryza
Fijivirus
biology.organism_classification
Plant Leaves
030104 developmental biology
Infectious Diseases
Biochemistry
Membrane protein
Agrobacterium tumefaciens
REOVIRIDAE
CIENCIAS AGRÍCOLAS
Host-Pathogen Interactions
Proteolysis
VIROPLASM
Agricultura
Silvicultura y Pesca
Sequence Alignment
CIENCIAS NATURALES Y EXACTAS
Protein Binding
Zdroj: Virus Research. 230:19-28
ISSN: 0168-1702
DOI: 10.1016/j.virusres.2017.01.002
Popis: Mal de Río Cuarto virus (MRCV) is a member of the Fijivirus genus, within the Reoviridae family, that replicates and assembles in cytoplasmic inclusion bodies called viroplasms. In this study, we investigated interactions between ten MRCV proteins by yeast two-hybrid (Y2H) assays and identified interactions of non-structural proteins P6/P6, P9-2/P9-2 and P6/P9-1. P9-1 and P6 are the major and minor components of the viroplasms respectively, whereas P9-2 is an N-glycosylated membrane protein of unknown function. Interactions involving P6 and P9-1 were confirmed by bimolecular fluorescence complementation (BiFC) in rice protoplasts. We demonstrated that a region including a predicted coiled-coil domain within the C-terminal moiety of P6 was necessary for P6/P6 and P6/P9-1 interactions. In turn, a short C-terminal arm was necessary for the previously reported P9-1 self-interaction. Transient expression of these proteins by agroinfiltration of Nicotiana benthamiana leaves showed very low accumulation levels and further in silico analyses allowed us to identify conserved PEST degradation sequences [rich in proline (P), glutamic acid (E), serine (S), and threonine (T)] within P6 and P9-1. The removal of these PEST sequences resulted in a significant increase of the accumulation of both proteins. Fil: Llauger, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: de Haro, Luis Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Alfonso, Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: del Vas, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina
Databáze: OpenAIRE
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