Regulation of adeno-associated virus gene expression in 293 cells: control of mRNA abundance and translation.
We studied the effects of the adeno-associated virus (AAV) rep gene on the control of gene expression from the AAV p40 promoter in 293 cells in the absence of an adenovirus coinfection. AAV vectors containing the chloramphenicol acetyltransferase (cat) gene were used to measure the levels of cat expression and steady-state mRNA from p40. When the rep gene was present in cis or in trans, cat expression from p40 was decreased 3- to 10-fold, but there was a 2- to 4-fold increase in the level of p40 mRNA. Conversely, cat expression increased and the p40 mRNA level decreased in the absence of the rep gene. Both wild-type and carboxyl-terminal truncated Rep proteins were capable of eliciting both effects. These data suggest two roles for the pleiotropic AAV rep gene: as a translational inhibitor and as a positive regulator of p40 mRNA levels. We also provide additional evidence for a cis-acting negative regulatory region which decreases RNA from the AAV p5 promoter in a fashion independent of rep.
Trempe
J P
JP
Laboratory of Molecular and Cellular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
Carter
B J
BJ
D000123:Acetyltransferases / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase; D000229:Dependovirus / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D014157:Transcription Factors / Q000502:physiology*; D014764:Viral Proteins / Q000502:physiology*
D012333:RNA, Messenger; D014157:Transcription Factors; D014764:Viral Proteins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase