Rev protein suppression of complex formation between nuclear proteins and rev-responsive element-containing RNA of human immunodeficiency virus-1

1995 ◽  
Vol 27 (12) ◽  
pp. 1317-1329 ◽  
Author(s):  
Andreas Bek ◽  
Werner E.G. Müller ◽  
Annie-Pierre Sève ◽  
Vadim Kavsan ◽  
Heinz C. Schröder
Keyword(s):  
Human Immunodeficiency Virus ◽  
Complex Formation ◽  
Nuclear Proteins ◽  
Immunodeficiency Virus ◽  
Responsive Element ◽  
Human Immunodeficiency Virus 1 ◽  
Rev Protein

scholarly journals Mutation of the Rev-binding loop in the human immunodeficiency virus 1 leader causes a replication defect characterized by altered RNA trafficking and packaging

2006 ◽  
Vol 87 (10) ◽  
pp. 3039-3044 ◽  
Author(s):  
Jane S. Greatorex ◽  
Elizabeth A. Palmer ◽  
Roger J. Pomerantz ◽  
John A. Dangerfield ◽  
Andrew M. L. Lever
Keyword(s):  
Human Immunodeficiency Virus ◽  
Nuclear Export ◽  
Rna Packaging ◽  
Packaging Signal ◽  
Virus Particles ◽  
Immunodeficiency Virus ◽  
Responsive Element ◽  
Viral Genomic Rna ◽  
Human Immunodeficiency Virus 1

An internal RNA loop, located within the packaging signal of human immunodeficiency virus 1, that resembles the Rev-responsive element (RRE) closely was identified previously. Subsequent in vitro studies confirmed that the loop, termed loop A, could bind Rev protein specifically. Its proximity to the major splice donor has suggested a role for Rev–loop A interaction supplementary to or preceding that of the Rev–RRE interaction. To investigate this further in a replication-competent provirus, loop A was mutated to decrease its affinity for Rev. Impairing the Rev–loop A interaction led to reduced nuclear export of viral genomic RNA. RNA packaging decreased by approximately 30 %. Viral protein production and export of virus particles appeared normal; however, the virus was severely replication-deficient. The loop A sequence, which is 98 % conserved amongst viral isolates, is implicated in several cis-acting functions critical to virus viability.

scholarly journals The human immunodeficiency virus type 1 Rev protein and the Rev-responsive element counteract the effect of an inhibitory 5' splice site in a 3' untranslated region.

1995 ◽  
Vol 15 (6) ◽  
pp. 2962-2971 ◽  
Author(s):  
S K Barksdale ◽  
C C Baker
Keyword(s):  
Gene Expression ◽  
Human Immunodeficiency Virus ◽  
Human Immunodeficiency Virus Type ◽  
Splice Site ◽  
Untranslated Region ◽  
Expression Vector ◽  
Immunodeficiency Virus ◽  
Responsive Element ◽  
Rev Protein

A 5' splice site located in a 3' untranslated region (3'UTR) has been shown previously to inhibit gene expression. Natural examples of inhibitory 5' splice sites have been identified in the late 3'UTRs of papillomaviruses and are thought to inhibit viral late gene expression at early stages of the viral life cycle. In this study, we demonstrate that the interaction of the human immunodeficiency virus type 1 Rev protein with the Rev-responsive element (RRE) overcomes the inhibitory effects of a 5' splice site located within a 3'UTR. This was studied by using both a bovine papillomavirus type 1 L1 cDNA expression vector and a chloramphenicol acetyltransferase expression vector containing a 5' splice site in the 3'UTR. In both systems, coexpression of Rev enhanced cytoplasmic expression from vectors containing the RRE even when the RRE and the inhibitory 5' splice site were separated by up to 1,000 nucleotides. In addition, multiple copies of a 5' splice site in a 3'UTR were shown to act synergistically, and this effect could also be moderated by the interaction of Rev and the RRE. These studies provide additional evidence that at least one mechanism of Rev action is through interactions with the splicing machinery. We have previously shown that base pairing between the U1 small nuclear RNA and a 3'UTR 5' splice site is required for inhibition of gene expression. However, experiments by J. Kjems and P. A. Sharp (J. Virol. 67:4769-4776, 1993) have suggested that Rev acts on spliceosome assembly at a stage after binding of the U1 small nuclear ribonucleoprotein to the 5' splice site. This finding suggests that binding of additional small nuclear ribonucleoproteins, as well as other splicing factors, may be necessary for the inhibitory action of a 3'UTR 5' splice site. These data also suggest that expression of the papillomavirus late genes in terminally differentiated keratinocytes can be regulated by a viral or cellular Rev-like activity.

A Clinical Trial of Retroviral-Mediated Transfer of arev-Responsive Element Decoy Gene Into CD34+Cells From the Bone Marrow of Human Immunodeficiency Virus-1–Infected Children

Blood ◽  
1999 ◽  
Vol 94 (1) ◽  
pp. 368-371 ◽  
Author(s):  
Donald B. Kohn ◽  
Gerhard Bauer ◽  
C. Robert Rice ◽  
J.C. Rothschild ◽  
Denise A. Carbonaro ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Bone Marrow ◽  
Human Immunodeficiency Virus ◽  
Gene Transfer ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Immunodeficiency Virus ◽  
Responsive Element ◽  
Hiv 1 ◽  
Human Immunodeficiency Virus 1

Genetic modification of hematopoietic stem cells with genes that inhibit replication of human immunodeficiency virus-1 (HIV-1) could lead to development of T lymphocytes and monocytic cells resistant to HIV-1 infection after transplantation. We performed a clinical trial to evaluate the safety and feasibility of this procedure, using bone marrow from four HIV-1–infected pediatric subjects (ages 8 to 17 years). We obtained bone marrow, isolated CD34+ cells, performed in vitro transduction with a retroviral vector carrying arev-responsive element (RRE) decoy gene, and reinfused the cells into these subjects with no evidence of adverse effects. The levels of gene-containing leukocytes in peripheral blood samples in the 1 year after gene transfer/cell infusion have been extremely low. These observations support the potential of performing gene therapy for HIV-1 using hematopoietic cells, but emphasize the need for improved gene transfer techniques.

scholarly journals Regulation of human immunodeficiency virus 1 transcription by nef microRNA

2005 ◽  
Vol 86 (3) ◽  
pp. 751-755 ◽  
Author(s):  
Shinya Omoto ◽  
Yoichi R. Fujii
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human T Cells ◽  
Immunodeficiency Virus ◽  
Infected Cells ◽  
Responsive Element ◽  
Transcriptional Pathway ◽  
Ltr Promoter ◽  
Hiv 1 ◽  
Human Immunodeficiency Virus 1

MicroRNAs (miRNAs) are ∼21–25 nt long and interact with mRNAs to lead to either translational repression or RNA cleavage through RNA interference. A previous study showed that human immunodeficiency virus 1 (HIV-1) nef dsRNA from AIDS patients who are long-term non-progressors inhibited HIV-1 transcription. In the study reported here, nef-derived miRNAs in HIV-1-infected and nef transduced cells were identified, and showed that HIV-1 transcription was suppressed by nef-expressing miRNA, miR-N367, in human T cells. The miR-N367 could reduce HIV-1 LTR promoter activity through the negative responsive element of the U3 region in the 5′-LTR. Therefore, nef miRNA produced in HIV-1-infected cells may downregulate HIV-1 transcription through both a post-transcriptional pathway and a transcriptional neo-pathway.

Inhibition of Formation of Rev-RRE Complex by Pyronin Y

1993 ◽  
Vol 4 (2) ◽  
pp. 103-111 ◽  
Author(s):  
H. C. Schröder ◽  
H. Ushijima ◽  
A. Bek ◽  
H. Merz ◽  
K. Pfeifer ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Complex Formation ◽  
Binding Studies ◽  
Immunodeficiency Virus ◽  
Responsive Element ◽  
Transcript Assembly ◽  
Hiv 1 ◽  
Rev Protein

The interaction of pyronin Y, an RNA intercalating drug, with the binding of Rev protein from human immunodeficiency virus type 1 (HIV-1) to Rev-responsive element (RRE)-containing env RNA was studied. In gel retardation assays, recombinant Rev protein tightly bound to in vitro transcribed RRE RNA. Nitrocellulose-filter-binding studies revealed a dissociation constant of ≈(1–2) = 10−10M (Pfeifer et al., 1991). Pyronin Y efficiently suppressed formation of the Rev-RRE complex. At a concentration of 1 μg ml−1, complex formation was almost completely inhibited. Electron microscopy showed that Rev oligomerizes in the presence of RRE-containing RNA with the formation of short rod-like structures or long filaments, depending on the length of the transcript. Assembly of Rev protein along RRE-containing RNAs was abolished after addition of pyronin Y. Thus pyronin Y represents the first compound described to inhibit Rev-RRE complex formation.

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