Analytical study of avian reticuloendotheliosis virus dimeric RNA generated in vivo and in vitro.
The retroviral genome consists of two identical RNA molecules associated at their 5' ends by a stable structure called the dimer linkage structure. The dimer linkage structure, while maintaining the dimer state of the retroviral genome, might also be involved in packaging and reverse transcription, as well as recombination during proviral DNA synthesis. To study the dimer structure of the retroviral genome and the mechanism of dimerization, we analyzed features of the dimeric genome of reticuloendotheliosis virus (REV) type A and identified elements required for its dimerization. Here we report that the REV dimeric genome extracted from virions and infected cells, as well as that synthesized in vitro, is more resistant to heat denaturation than avian sarcoma and leukemia virus, murine leukemia virus, or human immunodeficiency virus type 1 dimeric RNA. The minimal domain required to form a stable REV RNA dimer in vitro was found to map between positions 268 and 452 (KpnI and SalI sites), thus corresponding to the E encapsidation sequence (J. E. Embretson and H. M. Temin, J. Virol. 61:2675-2683, 1987). In addition, both the 5' and 3' halves of E are necessary in cis for RNA dimerization and the extent of RNA dimerization is influenced by viral sequences flanking E. Rapid and efficient dimerization of REV RNA containing gag sequences in addition to the E sequences and annealing of replication primer tRNA(Pro) to the primer-binding site necessitate the nucleocapsid protein.
Darlix
J L
JL
LaboRetro Institut National de la Santé et de la Recherche Medicale, Ecole Normale Supérieure de Lyon, France.
Gabus
C
C
Allain
B
B
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004285:Dogs; D004926:Escherichia coli / Q000235:genetics; D015750:Genes, gag; D016679:Genome, Viral; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D012361:RNA, Transfer, Pro / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D001356:Reticuloendotheliosis virus / Q000235:genetics* / Q000378:metabolism
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012361:RNA, Transfer, Pro; D012367:RNA, Viral