Characterization of the NSP6 protein product of rotavirus gene 11

• Published in: Virus research Vol. 130; no. 1; pp. 193 - 201
• Main Authors: Rainsford, Edward W., McCrae, Malcolm A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2007
• Subjects: Animals; Cell Line; Cercopithecus aethiops; Cytosol - chemistry; DNA, Viral - metabolism; Escherichia coli; Kinetics; Non-structural protein; RNA binding protein; RNA, Viral - metabolism; RNA-Binding Proteins - chemistry; RNA-Binding Proteins - genetics; RNA-Binding Proteins - isolation & purification; RNA-Binding Proteins - metabolism; Rotavirus; Rotavirus - physiology; Viral Nonstructural Proteins - chemistry; Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - isolation & purification; Viral Nonstructural Proteins - metabolism; Rotavirus; Non-structural protein; RNA binding protein
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/j.virusres.2007.06.011

The 12 kDa non-structural protein 6 (NSP6) is the least studied of the rotavirus proteins. In an attempt to further characterize this protein mono-specific antisera was generated using purified protein expressed in E. coli. Pulse/chase radio-labeling of virus infected cells was used to show that it is expressed at a steady but low rate throughout the virus replication cycle. In contrast to the other rotavirus non-structural proteins, NSP6 was found to have a high rate of turnover, being completely degraded within 2 h of synthesis. NSP6 tagged with GFP was used to probe the intracellular distribution of the protein, perinuclear aggregates were observed in the cytoplasm of transfected cells. Following virus infection of these transfected cells the aggregates were seen to redistribute to the viroplasms. Consistent with its localization to the site of viral genome replication and packaging, NSP6 was found to be a sequence independent nucleic acid binding protein, with similar affinities for ssRNA and dsRNA.