J. Theron et Lh. Nel, STABLE PROTEIN-RNA INTERACTION INVOLVES THE TERMINAL DOMAINS OF BLUETONGUE VIRUS MESSENGER-RNA, BUT NOT THE TERMINALLY CONSERVED SEQUENCES, Virology, 229(1), 1997, pp. 134-142
The interaction of bluetongue virus (BTV) proteins with viral RNA was
investigated in vitro by means of a biochemical approach. By subjectin
g cytoplasmic extracts from virus-infected baby hamster kidney cells a
nd in vitro synthesized radiolabeled RNA to ultraviolet cross-linking
assays, we demonstrated that, of all the BTV proteins, NS2 becomes mos
t intimately associated with the labeled viral RNA. Competition bindin
g studies indicated that NS2 has the greatest affinity for the 3' regi
on of the viral transcripts. By analyzing the binding efficiency of NS
2 to mutant RNA transcripts which lacked the fully conserved 5'- and/o
r 3'-terminal hexanucleotides, we have established that these sequence
s are not necessary for optimal binding, The specificity of the NS2-RN
A interaction was investigated by competition experiments with unlabel
ed BTV-specific homologous and heterologous competitor RNAs as well as
with viral double-stranded RNA (dsRNA). Although apparent differences
in the ability of NS2 to bind to the different RNA transcripts were o
bserved, it did not bind to the dsRNA. (C) 1997 Academic Press.