Cell Proteins Bind to A 67 Nucleotide Sequence within the 3’ Noncoding Region (NCR) of Simian Hemorrhagic Fever Virus (SFV) Negative-Strand RNA
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The 3’NCR of the SHFV negative-strand RNA [SHFV 3’(-)NCR RNA] is thought to be the initiation site of full-length and possibly also subgenomic positive-strand RNA and so is likely to contain cis-acting signals for viral RNA replication. Cellular and viral proteins may specifically interact with this region to form replication complexes. When in vitro transcribed SHFV 3’(-)NCR RNA was used as a probe in gel mobility shift assays, two RNA-protein complexes were detected with MA 104 S100 cytoplasmic extracts. The specificity of these RNA-protein interactions was demonstrated by competition gel mobil-ity shift assays. Four MA104 proteins (103, 86, 55, and 36 kDa) were detected by UV-in-duced cross-linking assays and three proteins (103, 55, and 36 kDa) were detected by northwestern blotting assays. The binding sites for these proteins were mapped to the re-gion between nucleotides 117 to 184 on the SHFV 3’(-)NCR RNA. Four cellular proteins with identical molecular masses to those of the proteins that bind to the SHFV 3’(-)NCR RNA were detected by the 3’(-) NCR of another arterivirus, LDV-C, suggesting that diver-gent arteriviruses utilize the same set of conserved cell protein domains.
KeywordsCellular Protein West Nile Virus Cytoplasmic Extract Identical Molecular Mass Simian Hemorrhagic Fever Virus
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