Mutation of the major 5' splice site renders a CMV-driven HIV-1 proviral clone Tat-dependent: connections between transcription and splicing.
Efficient transcription from the human immunodeficiency virus type 1 long terminal repeat (HIV-1 LTR) promoter is dependent on the viral transactivator Tat. To generate a Tat-independent proviral plasmid, we replaced the promoter in the HIV-1 LTR with the immediate early promoter of cytomegalovirus. Transfection of this plasmid yielded Tat-independent production of infectious HIV-1. Tat-independent expression was lost, however, when the major 5' splice site in the HIV-1 genome was mutated and no HIV-1-specific RNA or protein was detected. This defect was restored when a Tat expression plasmid was cotransfected. Our results support recent reports indicating an influence of the recognition of splice sites on efficient transcriptional elongation.
Bohne
Jens
J
Department of Virology, Universität Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany. bohne.jens@mh-hannover.de
Kräusslich
Hans-Georg
HG
D020121:5' Untranslated Regions; D017398:Alternative Splicing; D002999:Clone Cells; D003587:Cytomegalovirus / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D015696:Gene Products, tat / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000235:genetics*; D022821:RNA Splice Sites / Q000235:genetics*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D054322:tat Gene Products, Human Immunodeficiency Virus
D020121:5' Untranslated Regions; D015696:Gene Products, tat; D008164:Luminescent Proteins; D022821:RNA Splice Sites; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins