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Distinct RNA sequences in the gag region of human immunodeficiency virus type 1 decrease RNA stability and inhibit expression in the absence of Rev protein.

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  • 1. Human Retrovirus Section, National Cancer Institute-Frederick Cancer Research and Development Center, Maryland 21702-1201.
      Authors
    • Schwartz S 1
    (1 author)

Journal of Virology, 01 Jan 1992, 66(1):150-159
https://doi.org/10.1128/jvi.66.1.150-159.1992 PMID: 1727477 PMCID: PMC238270

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Abstract 

The expression of Gag, Pol, Vif, Vpr, Vpu, and Env proteins from unspliced and partially spliced human immunodeficiency virus type 1 (HIV-1) mRNAs depends on the viral protein Rev, while the production of Tat, Rev, and Nef from multiply spliced mRNAs does not require Rev. To investigate the difference between gag and tat mRNAs, we generated plasmids expressing tat-gag hybrid mRNAs. Insertion of the gag gene downstream of the tat open reading frame in the tat cDNA resulted in the inhibition of Tat production. This inhibition was caused, at least in part, by a decrease in the stability of the produced mRNA. Deletions in gag defined a 218-nucleotide inhibitory sequence named INS-1 and located at the 5' end of the gag gene. Further experiments indicated the presence of more than one inhibitory sequence in the gag-protease gene region of the viral genome. The inhibitory effect of INS-1 was counteracted by the positive effect mediated by the Rev-Rev-responsive element interaction, indicating that this sequence is important for Rev-regulated gag expression. The INS-1 sequence did not contain any known HIV-1 splice sites and acted independently of splicing. It was found to have an unusually high AU content (61.5% AU), a common feature among cellular mRNAs with short half-lives. These results suggest that HIV-1 and possibly other lentiviruses have evolved to express unstable mRNAs which require additional regulatory factors for their expression. This strategy may offer the virus several advantages, including the ability to enter a state of low or latent expression in the host.

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J Virol. 1992 Jan; 66(1): 150–159.
PMCID: PMC238270
PMID: 1727477

Distinct RNA sequences in the gag region of human immunodeficiency virus type 1 decrease RNA stability and inhibit expression in the absence of Rev protein.

S Schwartz, B K Felber, and G N Pavlakis
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Abstract

The expression of Gag, Pol, Vif, Vpr, Vpu, and Env proteins from unspliced and partially spliced human immunodeficiency virus type 1 (HIV-1) mRNAs depends on the viral protein Rev, while the production of Tat, Rev, and Nef from multiply spliced mRNAs does not require Rev. To investigate the difference between gag and tat mRNAs, we generated plasmids expressing tat-gag hybrid mRNAs. Insertion of the gag gene downstream of the tat open reading frame in the tat cDNA resulted in the inhibition of Tat production. This inhibition was caused, at least in part, by a decrease in the stability of the produced mRNA. Deletions in gag defined a 218-nucleotide inhibitory sequence named INS-1 and located at the 5' end of the gag gene. Further experiments indicated the presence of more than one inhibitory sequence in the gag-protease gene region of the viral genome. The inhibitory effect of INS-1 was counteracted by the positive effect mediated by the Rev-Rev-responsive element interaction, indicating that this sequence is important for Rev-regulated gag expression. The INS-1 sequence did not contain any known HIV-1 splice sites and acted independently of splicing. It was found to have an unusually high AU content (61.5% AU), a common feature among cellular mRNAs with short half-lives. These results suggest that HIV-1 and possibly other lentiviruses have evolved to express unstable mRNAs which require additional regulatory factors for their expression. This strategy may offer the virus several advantages, including the ability to enter a state of low or latent expression in the host.

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Selected References

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  • Ahmed YF, Hanly SM, Malim MH, Cullen BR, Greene WC. Structure-function analyses of the HTLV-I Rex and HIV-1 Rev RNA response elements: insights into the mechanism of Rex and Rev action. Genes Dev. 1990 Jun;4(6):1014–1022. [Abstract] [Google Scholar]
  • Arrigo SJ, Chen IS. Rev is necessary for translation but not cytoplasmic accumulation of HIV-1 vif, vpr, and env/vpu 2 RNAs. Genes Dev. 1991 May;5(5):808–819. [Abstract] [Google Scholar]
  • Arya SK, Gallo RC. Three novel genes of human T-lymphotropic virus type III: immune reactivity of their products with sera from acquired immune deficiency syndrome patients. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2209–2213. [Europe PMC free article] [Abstract] [Google Scholar]
  • Benko DM, Robinson R, Solomin L, Mellini M, Felber BK, Pavlakis GN. Binding of trans-dominant mutant Rev protein of human immunodeficiency virus type 1 to the cis-acting Rev-responsive element does not affect the fate of viral mRNA. New Biol. 1990 Dec;2(12):1111–1122. [Abstract] [Google Scholar]
  • Berger J, Hauber J, Hauber R, Geiger R, Cullen BR. Secreted placental alkaline phosphatase: a powerful new quantitative indicator of gene expression in eukaryotic cells. Gene. 1988 Jun 15;66(1):1–10. [Abstract] [Google Scholar]
  • Black AC, Chen IS, Arrigo S, Ruland CT, Allogiamento T, Chin E, Rosenblatt JD. Regulation of HTLV-II gene expression by Rex involves positive and negative cis-acting elements in the 5'long terminal repeat. Virology. 1991 Apr;181(2):433–444. [Abstract] [Google Scholar]
  • Brawerman G. Determinants of messenger RNA stability. Cell. 1987 Jan 16;48(1):5–6. [Abstract] [Google Scholar]
  • Chang DD, Sharp PA. Regulation by HIV Rev depends upon recognition of splice sites. Cell. 1989 Dec 1;59(5):789–795. [Abstract] [Google Scholar]
  • Clavel F, Orenstein JM. A mutant of human immunodeficiency virus with reduced RNA packaging and abnormal particle morphology. J Virol. 1990 Oct;64(10):5230–5234. [Europe PMC free article] [Abstract] [Google Scholar]
  • Cleveland DW, Yen TJ. Multiple determinants of eukaryotic mRNA stability. New Biol. 1989 Nov;1(2):121–126. [Abstract] [Google Scholar]
  • Cochrane AW, Chen CH, Rosen CA. Specific interaction of the human immunodeficiency virus Rev protein with a structured region in the env mRNA. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1198–1202. [Europe PMC free article] [Abstract] [Google Scholar]
  • Daefler S, Klotman ME, Wong-Staal F. Trans-activating rev protein of the human immunodeficiency virus 1 interacts directly and specifically with its target RNA. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4571–4575. [Europe PMC free article] [Abstract] [Google Scholar]
  • Daly TJ, Cook KS, Gray GS, Maione TE, Rusche JR. Specific binding of HIV-1 recombinant Rev protein to the Rev-responsive element in vitro. Nature. 1989 Dec 14;342(6251):816–819. [Abstract] [Google Scholar]
  • Dayton AI, Terwilliger EF, Potz J, Kowalski M, Sodroski JG, Haseltine WA. Cis-acting sequences responsive to the rev gene product of the human immunodeficiency virus. J Acquir Immune Defic Syndr. 1988;1(5):441–452. [Abstract] [Google Scholar]
  • Emerman M, Vazeux R, Peden K. The rev gene product of the human immunodeficiency virus affects envelope-specific RNA localization. Cell. 1989 Jun 30;57(7):1155–1165. [Abstract] [Google Scholar]
  • Feinberg MB, Jarrett RF, Aldovini A, Gallo RC, Wong-Staal F. HTLV-III expression and production involve complex regulation at the levels of splicing and translation of viral RNA. Cell. 1986 Sep 12;46(6):807–817. [Abstract] [Google Scholar]
  • Felber BK, Drysdale CM, Pavlakis GN. Feedback regulation of human immunodeficiency virus type 1 expression by the Rev protein. J Virol. 1990 Aug;64(8):3734–3741. [Europe PMC free article] [Abstract] [Google Scholar]
  • Felber BK, Hadzopoulou-Cladaras M, Cladaras C, Copeland T, Pavlakis GN. rev protein of human immunodeficiency virus type 1 affects the stability and transport of the viral mRNA. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1495–1499. [Europe PMC free article] [Abstract] [Google Scholar]
  • Felber BK, Paskalis H, Kleinman-Ewing C, Wong-Staal F, Pavlakis GN. The pX protein of HTLV-I is a transcriptional activator of its long terminal repeats. Science. 1985 Aug 16;229(4714):675–679. [Abstract] [Google Scholar]
  • Garrett ED, Tiley LS, Cullen BR. Rev activates expression of the human immunodeficiency virus type 1 vif and vpr gene products. J Virol. 1991 Mar;65(3):1653–1657. [Europe PMC free article] [Abstract] [Google Scholar]
  • Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. [Europe PMC free article] [Abstract] [Google Scholar]
  • Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. [Abstract] [Google Scholar]
  • Greenberg ME, Ziff EB. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. [Abstract] [Google Scholar]
  • Hadzopoulou-Cladaras M, Felber BK, Cladaras C, Athanassopoulos A, Tse A, Pavlakis GN. The rev (trs/art) protein of human immunodeficiency virus type 1 affects viral mRNA and protein expression via a cis-acting sequence in the env region. J Virol. 1989 Mar;63(3):1265–1274. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hammarskjöld ML, Heimer J, Hammarskjöld B, Sangwan I, Albert L, Rekosh D. Regulation of human immunodeficiency virus env expression by the rev gene product. J Virol. 1989 May;63(5):1959–1966. [Europe PMC free article] [Abstract] [Google Scholar]
  • Heaphy S, Dingwall C, Ernberg I, Gait MJ, Green SM, Karn J, Lowe AD, Singh M, Skinner MA. HIV-1 regulator of virion expression (Rev) protein binds to an RNA stem-loop structure located within the Rev response element region. Cell. 1990 Feb 23;60(4):685–693. [Abstract] [Google Scholar]
  • Jones TR, Cole MD. Rapid cytoplasmic turnover of c-myc mRNA: requirement of the 3' untranslated sequences. Mol Cell Biol. 1987 Dec;7(12):4513–4521. [Europe PMC free article] [Abstract] [Google Scholar]
  • Katz RA, Kotler M, Skalka AM. cis-acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implication for mechanisms of control. J Virol. 1988 Aug;62(8):2686–2695. [Europe PMC free article] [Abstract] [Google Scholar]
  • Katz RA, Skalka AM. Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Mol Cell Biol. 1990 Feb;10(2):696–704. [Europe PMC free article] [Abstract] [Google Scholar]
  • Knight DM, Flomerfelt FA, Ghrayeb J. Expression of the art/trs protein of HIV and study of its role in viral envelope synthesis. Science. 1987 May 15;236(4803):837–840. [Abstract] [Google Scholar]
  • Krawczyk Z, Wu C. Isolation of RNA for dot hybridization by heparin-DNase I treatment of whole cell lysate. Anal Biochem. 1987 Aug 15;165(1):20–27. [Abstract] [Google Scholar]
  • Lever A, Gottlinger H, Haseltine W, Sodroski J. Identification of a sequence required for efficient packaging of human immunodeficiency virus type 1 RNA into virions. J Virol. 1989 Sep;63(9):4085–4087. [Europe PMC free article] [Abstract] [Google Scholar]
  • Lu XB, Heimer J, Rekosh D, Hammarskjöld ML. U1 small nuclear RNA plays a direct role in the formation of a rev-regulated human immunodeficiency virus env mRNA that remains unspliced. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7598–7602. [Europe PMC free article] [Abstract] [Google Scholar]
  • Malim MH, Tiley LS, McCarn DF, Rusche JR, Hauber J, Cullen BR. HIV-1 structural gene expression requires binding of the Rev trans-activator to its RNA target sequence. Cell. 1990 Feb 23;60(4):675–683. [Abstract] [Google Scholar]
  • Malim MH, Hauber J, Le SY, Maizel JV, Cullen BR. The HIV-1 rev trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA. Nature. 1989 Mar 16;338(6212):254–257. [Abstract] [Google Scholar]
  • Morris-Vasios C, Kochan JP, Skalka AM. Avian sarcoma-leukosis virus pol-endo proteins expressed independently in mammalian cells accumulate in the nucleus but can be directed to other cellular compartments. J Virol. 1988 Jan;62(1):349–353. [Europe PMC free article] [Abstract] [Google Scholar]
  • Muesing MA, Smith DH, Cabradilla CD, Benton CV, Lasky LA, Capon DJ. Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. Nature. 1985 Feb 7;313(6002):450–458. [Abstract] [Google Scholar]
  • Olsen HS, Nelbock P, Cochrane AW, Rosen CA. Secondary structure is the major determinant for interaction of HIV rev protein with RNA. Science. 1990 Feb 16;247(4944):845–848. [Abstract] [Google Scholar]
  • Pavlakis GN, Felber BK. Regulation of expression of human immunodeficiency virus. New Biol. 1990 Jan;2(1):20–31. [Abstract] [Google Scholar]
  • Pomerantz RJ, Trono D, Feinberg MB, Baltimore D. Cells nonproductively infected with HIV-1 exhibit an aberrant pattern of viral RNA expression: a molecular model for latency. Cell. 1990 Jun 29;61(7):1271–1276. [Abstract] [Google Scholar]
  • Rabbitts PH, Forster A, Stinson MA, Rabbitts TH. Truncation of exon 1 from the c-myc gene results in prolonged c-myc mRNa stability. EMBO J. 1985 Dec 30;4(13B):3727–3733. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ratner L, Fisher A, Jagodzinski LL, Liou RS, Mitsuya H, Gallo RC, Wong-Staal F. Complete nucleotide sequences of functional clones of the virus associated with the acquired immunodeficiency syndrome, HTLV-III/LAV. Haematol Blood Transfus. 1987;31:404–406. [Abstract] [Google Scholar]
  • Ratner L, Fisher A, Jagodzinski LL, Mitsuya H, Liou RS, Gallo RC, Wong-Staal F. Complete nucleotide sequences of functional clones of the AIDS virus. AIDS Res Hum Retroviruses. 1987 Spring;3(1):57–69. [Abstract] [Google Scholar]
  • Rosen CA, Terwilliger E, Dayton A, Sodroski JG, Haseltine WA. Intragenic cis-acting art gene-responsive sequences of the human immunodeficiency virus. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2071–2075. [Europe PMC free article] [Abstract] [Google Scholar]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [Abstract] [Google Scholar]
  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [Abstract] [Google Scholar]
  • Schwartz S, Felber BK, Benko DM, Fenyö EM, Pavlakis GN. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):2519–2529. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schwartz S, Felber BK, Fenyö EM, Pavlakis GN. Env and Vpu proteins of human immunodeficiency virus type 1 are produced from multiple bicistronic mRNAs. J Virol. 1990 Nov;64(11):5448–5456. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schwartz S, Felber BK, Pavlakis GN. Expression of human immunodeficiency virus type 1 vif and vpr mRNAs is Rev-dependent and regulated by splicing. Virology. 1991 Aug;183(2):677–686. [Abstract] [Google Scholar]
  • Seiki M, Hikikoshi A, Yoshida M. The U5 sequence is a cis-acting repressive element for genomic RNA expression of human T cell leukemia virus type I. Virology. 1990 May;176(1):81–86. [Abstract] [Google Scholar]
  • Shaw G, Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. [Abstract] [Google Scholar]
  • Shyu AB, Belasco JG, Greenberg ME. Two distinct destabilizing elements in the c-fos message trigger deadenylation as a first step in rapid mRNA decay. Genes Dev. 1991 Feb;5(2):221–231. [Abstract] [Google Scholar]
  • Shyu AB, Greenberg ME, Belasco JG. The c-fos transcript is targeted for rapid decay by two distinct mRNA degradation pathways. Genes Dev. 1989 Jan;3(1):60–72. [Abstract] [Google Scholar]
  • Sodroski J, Goh WC, Rosen C, Dayton A, Terwilliger E, Haseltine W. A second post-transcriptional trans-activator gene required for HTLV-III replication. Nature. 1986 May 22;321(6068):412–417. [Abstract] [Google Scholar]
  • Solomin L, Felber BK, Pavlakis GN. Different sites of interaction for Rev, Tev, and Rex proteins within the Rev-responsive element of human immunodeficiency virus type 1. J Virol. 1990 Dec;64(12):6010–6017. [Europe PMC free article] [Abstract] [Google Scholar]
  • Stoltzfus CM, Fogarty SJ. Multiple regions in the Rous sarcoma virus src gene intron act in cis to affect the accumulation of unspliced RNA. J Virol. 1989 Apr;63(4):1669–1676. [Europe PMC free article] [Abstract] [Google Scholar]
  • Stoltzfus CM, Lorenzen SK, Berberich SL. Noncoding region between the env and src genes of Rous sarcoma virus influences splicing efficiency at the src gene 3' splice site. J Virol. 1987 Jan;61(1):177–184. [Europe PMC free article] [Abstract] [Google Scholar]
  • Treisman R. Transient accumulation of c-fos RNA following serum stimulation requires a conserved 5' element and c-fos 3' sequences. Cell. 1985 Oct;42(3):889–902. [Abstract] [Google Scholar]
  • Wills JW, Craven RC, Achacoso JA. Creation and expression of myristylated forms of Rous sarcoma virus gag protein in mammalian cells. J Virol. 1989 Oct;63(10):4331–4343. [Europe PMC free article] [Abstract] [Google Scholar]
  • Wilson T, Treisman R. Removal of poly(A) and consequent degradation of c-fos mRNA facilitated by 3' AU-rich sequences. Nature. 1988 Nov 24;336(6197):396–399. [Abstract] [Google Scholar]
  • Wisdom R, Lee W. The protein-coding region of c-myc mRNA contains a sequence that specifies rapid mRNA turnover and induction by protein synthesis inhibitors. Genes Dev. 1991 Feb;5(2):232–243. [Abstract] [Google Scholar]
  • Wright CM, Felber BK, Paskalis H, Pavlakis GN. Expression and characterization of the trans-activator of HTLV-III/LAV virus. Science. 1986 Nov 21;234(4779):988–992. [Abstract] [Google Scholar]
  • Zapp ML, Green MR. Sequence-specific RNA binding by the HIV-1 Rev protein. Nature. 1989 Dec 7;342(6250):714–716. [Abstract] [Google Scholar]
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