A study of the dimer formation of Rous sarcoma virus RNA and of its effect on viral protein synthesis in vitro.
The genetic material of all retroviruses examined so far is an RNA dimer where two identical RNA subunits are joined at their 5' ends by a structure named dimer linkage structure (DLS). Since the precise location and structure of the DLS as well as the mechanism and role(s) of RNA dimerization remain unclear, we analysed the dimerization process of Rous sarcoma virus (RSV) RNA. For this purpose we set up an in vitro model for RSV RNA dimerization. Using this model RSV RNA was shown to form dimeric molecules and this dimerization process was greatly activated by nucleocapsid protein (NCp12) of RSV. Furthermore, RSV RNA dimerization was performed in the presence of complementary 5'32P-DNA oligomers in order to probe the monomer and dimer forms of RSV RNA. Data indicated that the DLS of RSV RNA probably maps between positions 544-564 from the 5' end. In an attempt to define sequences needed for the dimerization of RSV RNA, deletion mutageneses were generated in the 5' 600 nt. The results showed that the dimer promoting sequences probably are located within positions 208-270 and 400-600 from the 5' end and hence possibly encompassing the cis-acting elements needed for the specific encapsidation of RSV genomic RNA. Also it is reported that synthesis of the polyprotein precursor Pr76gag is inhibited upon dimerization of RSV RNA. These results suggest that dimerization and encapsidation of genome length RSV RNA might be linked in the course of virion formation since they appear to be under the control of the same cis elements, E and DLS, and the trans-acting factor nucleocapsid protein NCp12.
Bieth
E
E
LaboRetro, Centre de Recherches de Biochimie et de Génétique Cellulaires du CNRS, Toulouse, France.
Gabus
C
C
Darlix
J L
JL
D001358:Avian Sarcoma Viruses / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D001483:Base Sequence; D002213:Capsid / Q000378:metabolism*; D004252:DNA Mutational Analysis; D015683:Gene Products, gag / Q000235:genetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D012191:Retroviridae Proteins / Q000096:biosynthesis*; D014758:Viral Core Proteins / Q000378:metabolism*
D015683:Gene Products, gag; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D012191:Retroviridae Proteins; D014758:Viral Core Proteins