Four nucleotides are the minimal requirement for RNA recognition by rotavirus non-structural protein NSP3.

AUTOR(ES)

Poncet, D

RESUMO

The interaction of the group A rotavirus non-structural protein NSP3 (NSP3A) with RNA has been studied in vitro. Using semi-purified NSP3A protein expressed by a recombinant baculovirus and in vitro synthesized RNA, we determined by UV cross-linking and gel retardation assays that NSP3A binds, in a sequence-specific manner, the consensus sequence (AUGUGACC) present on the 3' ends of all group A rotavirus mRNAs. Using short oligoribonucleotides, we established that the minimal RNA sequence required for binding of NSP3A is GACC. Modifications of the UGACC oligonucleotide sequence impaired binding of the protein to the RNA. Furthermore, the recombinant NSP3 protein from rotavirus group C showed specificity for the 3' end consensus sequence (AUGUGGCU) of only group C mRNAs. Sequence analysis of the NSP3 proteins did not reveal significant homologies with other RNA binding proteins, thus the NSP3 proteins of rotaviruses are the prototypes of a new kind of sequence-specific RNA binding protein.

Documentos Relacionados

• The cytoplasmic tail of NSP4, the endoplasmic reticulum-localized non-structural glycoprotein of rotavirus, contains distinct virus binding and coiled coil domains.
• The shift from low to high non-structural protein 1 expression in rotavirus-infected MA-104 cells
• Topology of the non-structural rotavirus receptor glycoprotein NS28 in the rough endoplasmic reticulum.
• Nucleotide sequence of reovirus genome segment S3, encoding non-structural protein sigma NS.
• Multimers Formed by the Rotavirus Nonstructural Protein NSP2 Bind to RNA and Have Nucleoside Triphosphatase Activity