Inhibition of HIV-1 Replication in Lymphocytes by Mutants of the Rev Cofactor eIF-5A

Bevec, Dorian; Jaksche, Herbert; Oft, Martin; Wöhl, Thorsten; Himmelspach, Michèle; Pacher, Alexandra; Schebesta, Michael; Koettnitz, Karl; Dobrovnik, Marika; Csonga, Robert; Lottspeich, Friedrich; Hauber, Joachim

doi:10.1126/science.271.5257.1858

Cited by 193 publications

(169 citation statements)

References 20 publications

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“…Additional effects of Rev are likely to occur at the level of facilitating viral RNA transport or translation. These transport effects could involve the specific interplay of other cellular factors, such as the nucleoporin-like Rab͞Rip proteins (49)(50)(51) or eIF 5A (52,53), with the leucine-rich activation domain of Rev (7), which functions as a nuclear export signal (54,55). Mutation of this leucine-rich domain results in the formation of potent dominant-negative inhibitors of Rev (56), which already have shown promise in early clinical trials (57).…”

Section: Discussionmentioning

confidence: 99%

HIV Rev-dependent binding of SF2/ASF to the Rev response element: Possible role in Rev-mediated inhibition of HIV RNA splicing

Powell

Amaral

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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Production of the structural and enzymatic proteins of type 1 human immunodeficiency virus (HIV-1) is controlled by the rev regulatory gene product. The 116-amino acid Rev protein acts by binding to the Rev response element (RRE), a complex RNA stem-loop structure located within the env gene of HIV. Rev exerts a series of posttranscriptional effects, including the inhibition of viral RNA splicing, the activation of nuclear export of incompletely spliced viral RNAs, and the enhancement of translation of RRE-containing RNAs. Our studies now demonstrate that at least one member of the SR family of splicing factors, SF2͞ASF, specifically binds to a subregion of the RRE in vitro in a Rev-dependent manner. Furthermore, expression of high levels of SF2͞ASF inhibits Rev function and impairs HIV replication in vivo. Both the in vitro binding of SF2͞ASF to the Rev͞RRE complex and the in vivo inhibition of Rev action by SF2͞ASF are abrogated by mutation of the N-terminal RNA recognition motif but are not affected by mutation of the C-terminal arginine-serinerich domain. These findings suggest that Rev inhibition of HIV splicing likely involves recruitment of the essential splicing factor SF2͞ASF to the Rev͞RRE complex. However, these inhibitory effects of Rev on viral RNA splicing are apparently overcome by augmenting the intracellular levels of SF2͞ASF expression.Rev is an essential regulatory protein of the type 1 human immunodeficiency virus (HIV-1) that controls production of proteins required for the assembly of infectious virions (1-5). The 9-kb full-length genomic transcript of HIV undergoes a complex series of posttranscriptional splicing events resulting in the generation of more than 20 mRNAs that encode 9 different viral proteins (for review see refs. 6-8). The resultant mRNAs can be grouped in three size classes: the 2-kb viral RNAs are fully spliced and encode the regulatory proteins of HIV, including Rev, Tat, and Nef, whereas the 9-and 4-kb mRNAs encode the structural, enzymatic, and ancillary viral proteins, including Gag, Pol, Vif, Vpu, Vpr, and Env. The appearance of the 4-and 9-kb classes of mRNAs in the cytoplasm is regulated by the Rev protein (1-5). Rev binds specifically to the Rev response element (RRE), a complex 220-nucleotide RNA stem-loop structure located in the env coding sequence. This binding leads to effective cytoplasmic expression of the singly spliced (4 kb) and unspliced (9 kb) viral mRNAs, thus allowing the transition from early-stage regulatory protein synthesis to late-stage structural and enzymatic viral protein production (2, 6, 7). In the absence of Rev, these incompletely spliced RRE-containing transcripts are retained in the nucleus where they are either completely spliced or degraded (6, 7). Various mechanisms have been suggested to explain the action of Rev, including inhibition of viral mRNA splicing, activation of viral RNA transport from the nucleus to the cytoplasm, and enhanced translation of RRE-containing viral RNAs (1, 6-8). These mechanisms are not mutually ex...

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Section: Discussionmentioning

confidence: 99%

HIV Rev-dependent binding of SF2/ASF to the Rev response element: Possible role in Rev-mediated inhibition of HIV RNA splicing

Powell

Amaral

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

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“…3G). According to previous data (33,45), these spots represent the unhypusinated (Fig. 3G, red arrow) and the more acidic unhypusinated and acetylated form of eIF-5A1 (Fig.…”

Section: Dhs Is Crucial For Embryonic Development As Well As For Viabmentioning

confidence: 99%

Biological Relevance and Therapeutic Potential of the Hypusine Modification System

Pällmann

Braig

Sievert

et al. 2015

Journal of Biological Chemistry

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Background: Hypusine modification of the eukaryotic initiation factor 5A (eIF-5A) represents a conserved post-translational modification that regulates translation. Results: Deletion of hypusine modification enzymes exerts strong phenotypes. eIF-5A2-deleted animals are viable and fertile. Conclusion: Both enzymatic steps of hypusine modification are essential for mammalian homeostasis, whereas the cancerrelated isoform eIF-5A2 is dispensable. Significance: eIF-5A2 might represent a safe therapeutic target.

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Section: Human Immunodeficiency Virus Rev Facilitates the Cytoplasmicmentioning

confidence: 99%

“…Rev's effector domain contains a leucine-rich nuclear export signal (NES), which binds the nuclear export receptor CRM1 (Weis 2002). Other cellular proteins have been shown to interact directly or indirectly with the Rev NES, including the kinesin-like Rev/Rex effector binding protein (REBP) and the eukaryotic initiation factor-5 alpha (eIF-5␣; Bevec et al 1996;Dayton 1996;Kjems and Askjaer 2000;Venkatesh et al 2003).The human Rev-interacting protein (hRIP), also known as hRab or Hrb, was identified using a yeast twohybrid screen with Rev as the bait (Bogerd et al 1995;Fritz et al 1995). Additional studies demonstrated a strong correlation between the ability of Rev NES mutants to support Rev function in yeast and their ability to interact with hRIP in the two-hybrid assay (Stutz et al 1996).…”

mentioning

confidence: 99%

hRIP, a cellular cofactor for Rev function, promotes release of HIV RNAs from the perinuclear region

Sánchez-Velar

Udofia²,

Yu³

et al. 2003

Genes Dev.

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Human immunodeficiency virus Rev facilitates the cytoplasmic accumulation of viral RNAs that contain aRev binding site. A human Rev-interacting protein (hRIP) was originally identified based on its ability to interact with the Rev nuclear export signal (NES) in yeast two-hybrid assays. To date, however, the function of hRIP and a role for hRIP in Rev-directed RNA export have remained elusive. Here we ablate hRIP activity with a dominant-negative mutant or RNA interference and analyze Rev function by RNA in situ hybridization. We find, unexpectedly, that in the absence of functional hRIP, Rev-directed RNAs mislocalize and aberrantly accumulate at the nuclear periphery, where hRIP is localized. In contrast, in the absence of Rev or the Rev cofactor CRM1, Rev-directed RNAs remain nuclear. We further show that the RNA mislocalization pattern resulting from loss of hRIP activity is highly specific to Rev function: the intracellular distribution of cellular poly(A) + mRNA, nuclear proteins, and, most important, NES-containing proteins, are unaffected. Thus, hRIP is an essential cellular Rev cofactor, which acts at a previously unanticipated step in HIV-1 RNA export: movement of RNAs from the nuclear periphery to the cytoplasm. Animal viruses often encode regulatory proteins, which facilitate expression of their viral genes in the host cell. One such example is the human immunodeficiency virus type 1 (HIV-1) Rev protein, which uses a novel mechanism to regulate viral messenger RNA (mRNA) export from the nucleus. Rev is a sequence-specific RNA binding protein that facilitates the cytoplasmic accumulation of the intron-containing HIV-1 gag-pol and env mRNAs (Cullen 2002). Rev interacts with a cis-acting viral RNA element in these mRNAs designated the Rev responsive element (RRE; Pollard and Malim 1998; Hope 1999). Rev contains two distinct functional domains: an arginine-rich RNA binding motif (ARM) and a leucinerich "effector" domain. Rev's ARM domain confers sequence-specific RNA binding, protein oligomerization, and nuclear targeting (Frankel and Young 1998). Rev's effector domain contains a leucine-rich nuclear export signal (NES), which binds the nuclear export receptor CRM1 (Weis 2002). Other cellular proteins have been shown to interact directly or indirectly with the Rev NES, including the kinesin-like Rev/Rex effector binding protein (REBP) and the eukaryotic initiation factor-5 alpha (eIF-5␣; Bevec et al. 1996;Dayton 1996;Kjems and Askjaer 2000;Venkatesh et al. 2003).The human Rev-interacting protein (hRIP), also known as hRab or Hrb, was identified using a yeast twohybrid screen with Rev as the bait (Bogerd et al. 1995;Fritz et al. 1995). Additional studies demonstrated a strong correlation between the ability of Rev NES mutants to support Rev function in yeast and their ability to interact with hRIP in the two-hybrid assay (Stutz et al. 1996). However, subsequent genetic analyses indicate the Rev-hRIP interaction is indirect, and is likely bridged by CRM1 (Fritz and Green 1996;Henderson and Percipalle 1997;Ne...

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Inhibition of HIV-1 Replication in Lymphocytes by Mutants of the Rev Cofactor eIF-5A

Cited by 193 publications

References 20 publications

HIV Rev-dependent binding of SF2/ASF to the Rev response element: Possible role in Rev-mediated inhibition of HIV RNA splicing

HIV Rev-dependent binding of SF2/ASF to the Rev response element: Possible role in Rev-mediated inhibition of HIV RNA splicing

Biological Relevance and Therapeutic Potential of the Hypusine Modification System

hRIP, a cellular cofactor for Rev function, promotes release of HIV RNAs from the perinuclear region

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