The cis-acting 3'-element of rubella virus RNA has DNA promoter activity.
The 3'-terminal region of the rubella virus (RV) positive-strand RNA, referred to here as the cis-acting element (CAE), is implicated in the initiation of negative-strand RNA synthesis. Sequence analysis of the 3'-CAE shows that there is a putative TATA box which is surrounded by G + C-rich sequences. To determine whether this element, in a DNA form, has the capability to initiate transcription, a 3'-end 165-bp NarI-EcoRI fragment from the RV cDNA was cloned upstream from a cat reporter gene. The level of CAT activity was dependent on the presence of the 3'-CAE and the SV40 enhancer. Primer extension analysis of the CAT mRNA showed that the transcription start point is in the RV 3'-CAE, 34 bp downstream from the putative TATA box. DNA-gel shift analysis revealed that three nucleoprotein-specific complexes were formed with the 3'-CAE and the binding sites for these proteins were between bp -64 to -108. The possible promoter function of the RV 3'-CAE is discussed in context to RV persistence.
Cao
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Division of Biochemistry and Biophysics, Food and Drug Administration, Bethesda, MD 20892.
Liu
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Nakhasi
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HL
D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000378:metabolism / Q000502:physiology; D004742:Enhancer Elements, Genetic; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012367:RNA, Viral / Q000502:physiology*; D012412:Rubella virus / Q000235:genetics*; D013539:Simian virus 40 / Q000235:genetics; D016385:TATA Box
D004279:DNA, Viral; D009687:Nuclear Proteins; D012367:RNA, Viral; D015500:Chloramphenicol O-Acetyltransferase