HIV-1 Rev can specifically interact with MMTV RNA and upregulate gene expression.
We present evidence that the HIV-1 Rev protein can heterologously regulate expression of the simple beta retrovirus mouse mammary tumour virus (MMTV). Up to 10-fold upregulation was seen in a functional assay system when specific MMTV sequences were substituted for the HIV-1 Rev responsive element (RRE). RNA gel shift analysis showed that purified recombinant Rev could specifically bind to MMTV unique region 3 prime (U3) RNA and that these sequences could compete for wild-type Rev-RRE binding approximately 20-fold more efficiently than a non-specific competitor RNA. Using a combination of in silico and deletion mutation analyses, it was not possible to define any single specific secondary structure responsive to Rev, suggesting that a structure or combination of structures that only form in the context of the complete U3 transcript is/are required to interact with Rev. Taken together, these results suggest that HIV-1 Rev can directly bind to MMTV RNA as well as mediate upregulation of MMTV gene expression.
Dangerfield
John A
JA
Research Institute of Virology and Biomedicine, University of Veterinary Medicine, Vienna, Austria. john.dangerfield@vu-wien.ac.at
Hohenadl
Christine
C
Egerbacher
Monika
M
Kodajova
Petra
P
Salmons
Brian
B
Günzburg
Walter H
WH
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D024202:Electrophoretic Mobility Shift Assay; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology; D015854:Up-Regulation / Q000502:physiology*; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus