Control of retroviral RNA splicing through maintenance of suboptimal processing signals.
The full-length retroviral transcript serves as genomic RNA for progeny virions, as an mRNA for structural proteins and enzymes, and as a pre-mRNA substrate for splicing that yields subgenomic mRNAs that encode other essential proteins. Thus, RNA splicing to form subgenomic mRNAs must be incomplete or regulated in order to preserve some of the full-length transcripts. We have used the avian sarcoma virus system to delineate the viral functions that are required in the regulation of the splicing event that forms the envelope glycoprotein (env) subgenomic mRNA. We observed previously that a specific insertion mutation just 5' of the env splice acceptor site resulted in nearly complete splicing to form env mRNA and a concomitant replication defect which is presumably due to a deficit of the full-length transcript. Replication-competent pseudorevertants contained second-site mutations that restored splicing control, and these mapped either just upstream or downstream of the env splice acceptor site. In this report, we show that splicing control at this site does not require expression of any known viral replication protein(s), nor does it appear to require the viral splice donor site. From these results and analysis of additional splicing mutations obtained by in vivo selection, we conclude that splicing is controlled through the maintenance of suboptimal cis-acting signals in the viral RNA that alter the efficiency of recognition by the cellular splicing machinery.
Katz
R A
RA
Fox Chase Cancer Center, Institute for Cancer Research, Philadelphia, Pennsylvania 19111.
Skalka
A M
AM
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D005091:Exons; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics; D014771:Virion / Q000235:genetics; D014779:Virus Replication
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins