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PMID: 7966585 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

Continuous propagation of RRE(-) and Rev(-)RRE(-) human immunodeficiency virus type 1 molecular clones containing a cis-acting element of simian retrovirus type 1 in human peripheral blood lymphocytes.

Journal of virology ·Vol. 68 ·No. 12 ·1994-12-00 ·Pages 7944-52

Zolotukhin AS, Valentin A, Pavlakis GN, Felber BK

Abstract

Molecular clones of human immunodeficiency virus type 1 that contained either 37 point mutations in the Rev-responsive element (RRE) that did not affect the overlapping env reading frame or both a mutated RRE and two mutations that eliminated Rev were constructed. The mutations in the RRE were shown to remove both negative and Rev-inducible positive effects of the RRE on gene expression (G. Nasioulas, A. S. Zolotukhin, C. Tabernero, L. Solomin, C. P. Cunningham, G. N. Pavlakis, and B. K. Felber, J. Virol. 68:2986-2993, 1994). Upon insertion of a cis-acting element of simian retrovirus type 1 (SRV-1) into these clones, both RRE(-) and Rev(-)RRE(-) clones were expressed efficiently. The element of SRV-1 has properties similar to those of the recently identified element of Mason-Pfizer monkey virus (M. Bray, S. Prasad, J. W. Dubay, E. Hunter, K.-T. Jeang, D. Rekosh, and M.-L. Hammarskjold, Proc. Natl. Acad. Sci. USA 4:1256-1260, 1994). We demonstrated that virus preparations produced after transfections of these SRV-1 element-containing molecular clones in human cells were infectious after cell-free transmission, that they replicated about 5 to 10 times less efficiently than wild-type virus, and that they were propagated continuously for more than 7 months in human peripheral blood mononuclear cells. Growth characteristics and sequence analysis of these viruses after long-term culture demonstrated that no RRE(+)Rev(+) revertants developed. These data demonstrate that human immunodeficiency virus type 1 Rev and RRE can be replaced by heterologous regulatory systems, resulting in efficient virus production. The resulting Rev(-)RRE(-) virus can be prepared and propagated efficiently in tissue culture and can be used for further studies of the life cycle of the virus. The data also suggest that Rev acts exclusively through the RRE interaction and that it does not have any additional essential function in the life cycle of the virus.

Related Genes
env nef rev tat vif vpu
MeSH Terms
Base Sequence Cell Line Cells, Cultured Chromosome Mapping Cloning, Molecular DNA Primers DNA, Viral/chemistry,genetics Gene Products, rev/metabolism Genes, env HIV-1/genetics,physiology HeLa Cells Humans Kinetics Lymphocytes/virology Molecular Sequence Data Mutagenesis, Site-Directed Open Reading Frames Point Mutation Proviruses/genetics,physiology Retroviruses, Simian/genetics Sequence Homology, Nucleic Acid Time Factors Transfection Virus Replication rev Gene Products, Human Immunodeficiency Virus
Chemicals
DNA Primers DNA, Viral Gene Products, rev rev Gene Products, Human Immunodeficiency Virus
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zolotukhin A S
Human Retrovirus Pathogenesis Group, National Cancer Institute--Frederick Cancer Research and Development Center, Maryland 21702-1201.
Valentin A
Pavlakis G N
Felber B K
References (47)
47 references, click to expand
  1. Comparative analysis of the HTLV-I Rex and HIV-1 Rev trans-regulatory proteins and their RNA response elements.
    Genes Dev. 1989 Oct;3(10):1534-44 PMID: 2482226
  2. Functional analysis of CAR, the target sequence for the Rev protein of HIV-1.
    Science. 1989 Dec 22;246(4937):1625-9 PMID: 2688093
  3. Secondary structure is the major determinant for interaction of HIV rev protein with RNA.
    Science. 1990 Feb 16;247(4944):845-8 PMID: 2406903
  4. Identification of a cis-acting element in human immunodeficiency virus type 2 (HIV-2) that is responsive to the HIV-1 rev and human T-cell leukemia virus types I and II rex proteins.
    J Virol. 1990 Apr;64(4):1690-7 PMID: 2157051
  5. A novel human immunodeficiency virus type 1 protein, tev, shares sequences with tat, env, and rev proteins.
    J Virol. 1990 Jun;64(6):2505-18 PMID: 2186172
  6. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1.
    J Virol. 1990 Jun;64(6):2519-29 PMID: 2335812
  7. Regulation of HIV-1 gene expression.
    Annu Rev Immunol. 1990;8:453-75 PMID: 2188670
  8. Simple, sensitive, and specific detection of human immunodeficiency virus type 1 in clinical specimens by polymerase chain reaction with nested primers.
    J Clin Microbiol. 1990 Jul;28(7):1560-4 PMID: 2380380
  9. 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):1014-22 PMID: 2116986
  10. Tat and Rev: positive regulators of HIV gene expression.
    AIDS. 1990 Jun;4(6):499-509 PMID: 2201316
  11. Human immunodeficiency virus rev protein recognizes a target sequence in rev-responsive element RNA within the context of RNA secondary structure.
    J Virol. 1990 Dec;64(12):5966-75 PMID: 2243382
  12. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  13. A molecular clone of HTLV-III with biological activity.
    Nature. 1985 Jul 18-24;316(6025):262-5 PMID: 2410792
  14. Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone.
    J Virol. 1986 Aug;59(2):284-91 PMID: 3016298
  15. Firefly luciferase gene: structure and expression in mammalian cells.
    Mol Cell Biol. 1987 Feb;7(2):725-37 PMID: 3821727
  16. Sequence relationships of type D retroviruses which cause simian acquired immunodeficiency syndrome.
    Virology. 1987 Apr;157(2):317-29 PMID: 2435057
  17. Functional replacement of the HIV-1 rev protein by the HTLV-1 rex protein.
    Nature. 1988 Oct 20;335(6192):738-40 PMID: 3262832
  18. 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):1265-74 PMID: 2783738
  19. 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):6010-7 PMID: 2243384
  20. Cross-activation of the Rex proteins of HTLV-I and BLV and of the Rev protein of HIV-1 and nonreciprocal interactions with their RNA responsive elements.
    New Biol. 1989 Dec;1(3):318-28 PMID: 2562124
  21. Structural analysis of the interaction between the human immunodeficiency virus Rev protein and the Rev response element.
    Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):683-7 PMID: 1992459
  22. Human immunodeficiency virus as a prototypic complex retrovirus.
    J Virol. 1991 Mar;65(3):1053-6 PMID: 1995941
  23. 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):1111-22 PMID: 2088501
  24. 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):808-19 PMID: 1827422
  25. Characterization of HIV-1 REV protein: binding stoichiometry and minimal RNA substrate.
    Nucleic Acids Res. 1991 Apr 11;19(7):1577-83 PMID: 2027765
  26. The type I human T-cell leukemia virus (HTLV-I) Rex trans-activator binds directly to the HTLV-I Rex and the type 1 human immunodeficiency virus Rev RNA response elements.
    Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5704-8 PMID: 1905815
  27. The Rex regulatory protein of human T-cell lymphotropic virus type I binds specifically to its target site within the viral RNA.
    Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7145-9 PMID: 1871127
  28. Human immunodeficiency virus type 1 regulator of virion expression, rev, forms nucleoprotein filaments after binding to a purine-rich "bubble" located within the rev-responsive region of viral mRNAs.
    Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7366-70 PMID: 1871141
  29. The biochemistry of AIDS.
    Annu Rev Biochem. 1991;60:577-630 PMID: 1883204
  30. HIV-1 Rev regulation involves recognition of non-Watson-Crick base pairs in viral RNA.
    Cell. 1991 Nov 1;67(3):529-36 PMID: 1934059
  31. Extensive sequence-specific information throughout the CAR/RRE, the target sequence of the human immunodeficiency virus type 1 Rev protein.
    J Virol. 1992 Feb;66(2):1139-51 PMID: 1731093
  32. The Rev protein of human immunodeficiency virus type 1 promotes polysomal association and translation of gag/pol and vpu/env mRNAs.
    Mol Cell Biol. 1992 Mar;12(3):1375-86 PMID: 1545819
  33. The HIV-1 Rev protein: a model system for coupled RNA transport and translation.
    New Biol. 1991 Dec;3(12):1220-32 PMID: 1725960
  34. A specific sequence with a bulged guanosine residue(s) in a stem-bulge-stem structure of Rev-responsive element RNA is required for trans activation by human immunodeficiency virus type 1 Rev.
    J Virol. 1992 Jun;66(6):3699-706 PMID: 1583728
  35. Mutational inactivation of an inhibitory sequence in human immunodeficiency virus type 1 results in Rev-independent gag expression.
    J Virol. 1992 Dec;66(12):7176-82 PMID: 1433510
  36. Protective effects of a live attenuated SIV vaccine with a deletion in the nef gene.
    Science. 1992 Dec 18;258(5090):1938-41 PMID: 1470917
  37. A proposal for a new approach to a preventive vaccine against human immunodeficiency virus type 1.
    Proc Natl Acad Sci U S A. 1993 May 15;90(10):4419-20 PMID: 8506281
  38. Packaging of human immunodeficiency virus type 1 RNA requires cis-acting sequences outside the 5' leader region.
    J Virol. 1993 Jul;67(7):3997-4005 PMID: 8510213
  39. Molecular biology of HIV-1: positive and negative regulatory elements important for virus expression.
    AIDS. 1993;7 Suppl 1:S51-62 PMID: 8363803
  40. A small element from the Mason-Pfizer monkey virus genome makes human immunodeficiency virus type 1 expression and replication Rev-independent.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1256-60 PMID: 8108397
  41. Elements distinct from human immunodeficiency virus type 1 splice sites are responsible for the Rev dependence of env mRNA.
    J Virol. 1994 May;68(5):2986-93 PMID: 8151769
  42. HIV with multiple gene deletions as a live attenuated vaccine for AIDS.
    AIDS Res Hum Retroviruses. 1992 Mar;8(3):411-21 PMID: 1571200
  43. 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):254-7 PMID: 2784194
  44. 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):1495-9 PMID: 2784208
  45. Regulation of human immunodeficiency virus env expression by the rev gene product.
    J Virol. 1989 May;63(5):1959-66 PMID: 2704072
  46. HTLV-1 rex and HIV-1 rev act through similar mechanisms to relieve suppression of unspliced RNA expression.
    Oncogene. 1989 Nov;4(11):1275-9 PMID: 2682457
  47. 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):685-93 PMID: 1689218
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.12.7944-7952.1994
Published
1994-12-00
Pages
7944-52
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237257
Subset
IM
Grants
NCI NIH HHS · N01-CO-74101 · United States
Databases
GENBANK
S74214
External Links
PubMed source DOI: 10.1128/JVI.68.12.7944-7952.1994 PMC Full Text View journal details
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