Home LiteratureArticle Details
PMID: 7966578 Published · ppublish English Comparative Study Journal Article

An integration-defective U5 deletion mutant of human immunodeficiency virus type 1 reverts by eliminating additional long terminal repeat sequences.

Journal of virology ·Vol. 68 ·No. 12 ·1994-12-00 ·Pages 7879-90

Vicenzi E, Dimitrov DS, Engelman A, Migone TS, Purcell DF, Leonard J, Englund G, Martin MA

Abstract

Nonoverlapping deletions that eliminated the 5' (HIV-1US/603del), middle (HIV-1U5/206del), and 3' (HIV-1U5/604del) thirds of the U5 region of the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) were studied for their effects on virus replication (transient transfection of HeLa cells) and infectivity (T-cell lines and peripheral blood mononuclear cells). All three mutants exhibited a wild-type phenotype in directing the production and release of virus particles from transfected HeLa cells. In infectivity assays, HIV-1U5/206del was usually indistinguishable from wild-type virus whereas HIV-1U%/603del was unable to infect human peripheral blood mononuclear cells or MT4 and CEM cells. Investigations of HIV-1U5/603del particles revealed a packaging defect resulting in a 10-fold reduction of encapsidated genomic RNA. The HIV-1U5/604del mutant either was noninfectious or exhibited delayed infection kinetics, depending on the cell type and multiplicity of infection. Quantitative competitive PCR indicated that HIV-1U5/604del synthesized normal amounts of viral DNA in newly infected cells. During the course of a long-term infectivity assay, a revertant of the HIV-1U5/604del mutant that displayed rapid infection kinetics emerged. Nucleotide sequence analysis indicated that the original 26-nucleotide deletion present in HIV-1U5/604del had been extended an additional 19 nucleotides in the revertant virus. Characterization of the HIV-1U5/604del mutant LTR in in vitro integration reactions revealed defective 3' processing and strand transfer activities that were partially restored when the revertant LTR substrate was used, suggesting that the reversion corrected a similar defect in the mutant virus.

MeSH Terms
Base Sequence Cell Line Cells, Cultured DNA Primers Defective Viruses/genetics,physiology HIV Long Terminal Repeat HIV-1/genetics,physiology HeLa Cells Humans Lymphocytes/virology Molecular Sequence Data Oligodeoxyribonucleotides Polymerase Chain Reaction Proviruses/genetics,physiology RNA, Viral/biosynthesis Sequence Deletion Species Specificity Transfection Virus Integration Virus Replication
Chemicals
DNA Primers Oligodeoxyribonucleotides RNA, Viral
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Vicenzi E
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, National Cancer Institute, Bethesda, Maryland 20892.
Dimitrov D S
Engelman A
Migone T S
Purcell D F
Leonard J
Englund G
Martin M A
References (58)
58 references, click to expand
  1. Human immunodeficiency virus integrase protein requires a subterminal position of its viral DNA recognition sequence for efficient cleavage.
    J Virol. 1991 Sep;65(9):4636-44 PMID: 1870194
  2. Effect of internal viral sequences on the utility of retroviral vectors.
    J Virol. 1987 May;61(5):1647-50 PMID: 3033290
  3. HIV-1 DNA integration: mechanism of viral DNA cleavage and DNA strand transfer.
    Cell. 1991 Dec 20;67(6):1211-21 PMID: 1760846
  4. Both substrate and target oligonucleotide sequences affect in vitro integration mediated by human immunodeficiency virus type 1 integrase protein produced in Saccharomyces cerevisiae.
    J Virol. 1992 Apr;66(4):2359-68 PMID: 1548767
  5. Interaction between retroviral U5 RNA and the T psi C loop of the tRNA(Trp) primer is required for efficient initiation of reverse transcription.
    J Virol. 1992 Apr;66(4):2464-72 PMID: 1548772
  6. Changes in both gp120 and gp41 can account for increased growth potential and expanded host range of human immunodeficiency virus type 1.
    J Virol. 1992 Jul;66(7):4445-51 PMID: 1602552
  7. A mutation at one end of Moloney murine leukemia virus DNA blocks cleavage of both ends by the viral integrase in vivo.
    J Virol. 1992 Aug;66(8):5092-5 PMID: 1629963
  8. Identification of conserved amino acid residues critical for human immunodeficiency virus type 1 integrase function in vitro.
    J Virol. 1992 Nov;66(11):6361-9 PMID: 1404595
  9. Avian retroviral RNA encapsidation: reexamination of functional 5' RNA sequences and the role of nucleocapsid Cys-His motifs.
    J Virol. 1993 Jan;67(1):178-88 PMID: 8380070
  10. Quantitation of human immunodeficiency virus type 1 infection kinetics.
    J Virol. 1993 Apr;67(4):2182-90 PMID: 8445728
  11. Identification of discrete functional domains of HIV-1 integrase and their organization within an active multimeric complex.
    EMBO J. 1993 Aug;12(8):3269-75 PMID: 8344264
  12. Amino acid changes in the fourth conserved region of human immunodeficiency virus type 2 strain HIV-2ROD envelope glycoprotein modulate fusion.
    J Virol. 1993 Oct;67(10):6253-8 PMID: 8371358
  13. Alternative splicing of human immunodeficiency virus type 1 mRNA modulates viral protein expression, replication, and infectivity.
    J Virol. 1993 Nov;67(11):6365-78 PMID: 8411338
  14. Mutations in human immunodeficiency virus type 1 gp41 affect sensitivity to neutralization by gp120 antibodies.
    J Virol. 1993 Nov;67(11):6897-902 PMID: 8411395
  15. Modified nucleotides of tRNA(3Lys) modulate primer/template loop-loop interaction in the initiation complex of HIV-1 reverse transcription.
    J Biol Chem. 1993 Dec 5;268(34):25269-72 PMID: 7503978
  16. Increase in soluble CD4 binding to and CD4-induced dissociation of gp120 from virions correlates with infectivity of human immunodeficiency virus type 1.
    J Virol. 1994 Feb;68(2):1029-39 PMID: 7904656
  17. Selective extraction of polyoma DNA from infected mouse cell cultures.
    J Mol Biol. 1967 Jun 14;26(2):365-9 PMID: 4291934
  18. Heteroduplex analysis of avian RNA tumor viruses.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):477-81 PMID: 191811
  19. Terminal redundancy and the origin of replication of Rous sarcoma virus RNA.
    Proc Natl Acad Sci U S A. 1977 May;74(5):1908-12 PMID: 68472
  20. Terminally repeated sequences in the avian sarcoma virus RNA genome.
    Proc Natl Acad Sci U S A. 1977 Jun;74(6):2389-93 PMID: 70037
  21. Avian myeloblastosis virus RNA is terminally redundant: implications for the mechanism of retrovirus replication.
    Cell. 1977 Sep;12(1):57-72 PMID: 198142
  22. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  23. Function of the retrovirus long terminal repeat.
    Cell. 1982 Jan;28(1):3-5 PMID: 7066985
  24. Avian sarcoma and leukosis virus pol-endonuclease recognition of the tandem long terminal repeat junction: minimum site required for cleavage is also required for viral growth.
    J Virol. 1987 Jun;61(6):1999-2008 PMID: 3033327
  25. In vitro mutagenesis identifies a region within the envelope gene of the human immunodeficiency virus that is critical for infectivity.
    J Virol. 1988 Jan;62(1):139-47 PMID: 3257102
  26. A novel gene of HIV-1, vpu, and its 16-kilodalton product.
    Science. 1988 Sep 2;241(4870):1221-3 PMID: 3261888
  27. A retroviral RNA secondary structure required for efficient initiation of reverse transcription.
    J Virol. 1988 Oct;62(10):3622-30 PMID: 2458484
  28. Identification of a signal in a murine retrovirus that is sufficient for packaging of nonretroviral RNA into virions.
    J Virol. 1988 Oct;62(10):3802-6 PMID: 3418786
  29. Construction and analysis of deletion mutations in the U5 region of Moloney murine leukemia virus: effects on RNA packaging and reverse transcription.
    J Virol. 1989 Jan;63(1):319-27 PMID: 2908924
  30. Identification of a U5-specific sequence required for efficient polyadenylation within the human immunodeficiency virus long terminal repeat.
    J Virol. 1989 Jan;63(1):421-4 PMID: 2908926
  31. Integration of mini-retroviral DNA: a cell-free reaction for biochemical analysis of retroviral integration.
    Proc Natl Acad Sci U S A. 1989 May;86(9):3065-9 PMID: 2524066
  32. Structure of the termini of DNA intermediates in the integration of retroviral DNA: dependence on IN function and terminal DNA sequence.
    Cell. 1989 Jul 14;58(1):47-54 PMID: 2546673
  33. The NF-kappa B binding sites in the human immunodeficiency virus type 1 long terminal repeat are not required for virus infectivity.
    J Virol. 1989 Nov;63(11):4919-24 PMID: 2795721
  34. Mutations of RNA and protein sequences involved in human immunodeficiency virus type 1 packaging result in production of noninfectious virus.
    J Virol. 1990 May;64(5):1920-6 PMID: 2109098
  35. The IN protein of Moloney murine leukemia virus processes the viral DNA ends and accomplishes their integration in vitro.
    Cell. 1990 Aug 24;62(4):829-37 PMID: 2167180
  36. A mutant of human immunodeficiency virus with reduced RNA packaging and abnormal particle morphology.
    J Virol. 1990 Oct;64(10):5230-4 PMID: 2204725
  37. Generation of neutralization-resistant HIV-1 in vitro due to amino acid interchanges of third hypervariable env region.
    J Immunol. 1990 Nov 15;145(10):3240-6 PMID: 1700002
  38. Importance of the nef gene for maintenance of high virus loads and for development of AIDS.
    Cell. 1991 May 17;65(4):651-62 PMID: 2032289
  39. Overlapping retrovirus U5 sequence elements are required for efficient integration and initiation of reverse transcription.
    J Virol. 1991 Jul;65(7):3864-72 PMID: 1710292
  40. Contribution of NF-kappa B and Sp1 binding motifs to the replicative capacity of human immunodeficiency virus type 1: distinct patterns of viral growth are determined by T-cell types.
    J Virol. 1991 Aug;65(8):4350-8 PMID: 2072454
  41. Deletions in the tRNA(Lys) primer-binding site of human immunodeficiency virus type 1 identify essential regions for reverse transcription.
    J Virol. 1991 Sep;65(9):4555-64 PMID: 1714513
  42. A specific orientation of RNA secondary structures is required for initiation of reverse transcription.
    J Virol. 1994 Feb;68(2):611-8 PMID: 7507181
  43. Characterization of mutants of human immunodeficiency virus type 1 that have escaped neutralization by a monoclonal antibody to the gp120 V2 loop.
    J Virol. 1994 Feb;68(2):974-8 PMID: 7507188
  44. Evidence for a functional interaction between the V1/V2 and C4 domains of human immunodeficiency virus type 1 envelope glycoprotein gp120.
    J Virol. 1994 Apr;68(4):2503-12 PMID: 8139032
  45. Encapsidation sequences for spleen necrosis virus, an avian retrovirus, are between the 5' long terminal repeat and the start of the gag gene.
    Proc Natl Acad Sci U S A. 1982 Oct;79(19):5986-90 PMID: 6310558
  46. cis-Acting RNA packaging locus in the 115-nucleotide direct repeat of Rous sarcoma virus.
    J Virol. 1983 Dec;48(3):667-75 PMID: 6313966
  47. The terminal nucleotides of retrovirus DNA are required for integration but not virus production.
    Nature. 1983 Nov 10-16;306(5939):155-60 PMID: 6316141
  48. Construction and analysis of deletion mutations in the pol gene of Moloney murine leukemia virus: a new viral function required for productive infection.
    Cell. 1984 Jul;37(3):1043-52 PMID: 6204767
  49. A mutant murine leukemia virus with a single missense codon in pol is defective in a function affecting integration.
    Proc Natl Acad Sci U S A. 1984 Oct;81(20):6461-5 PMID: 6208550
  50. The retrovirus pol gene encodes a product required for DNA integration: identification of a retrovirus int locus.
    Proc Natl Acad Sci U S A. 1984 Dec;81(24):7885-9 PMID: 6083562
  51. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 PMID: 3881765
  52. Characterization of a continuous T-cell line susceptible to the cytopathic effects of the acquired immunodeficiency syndrome (AIDS)-associated retrovirus.
    Proc Natl Acad Sci U S A. 1985 Jul;82(13):4539-43 PMID: 2989831
  53. Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay.
    Science. 1985 Aug 9;229(4713):563-6 PMID: 2992081
  54. Mutants and pseudorevertants of Moloney murine leukemia virus with alterations at the integration site.
    Cell. 1985 Sep;42(2):573-80 PMID: 4028161
  55. A conserved cis-acting sequence in the 5' leader of avian sarcoma virus RNA is required for packaging.
    J Virol. 1986 Jul;59(1):163-7 PMID: 3012114
  56. 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
  57. Evidence that the packaging signal of Moloney murine leukemia virus extends into the gag region.
    J Virol. 1987 May;61(5):1639-46 PMID: 3502707
  58. Substrate specificity of recombinant human immunodeficiency virus integrase protein.
    J Virol. 1991 Oct;65(10):5624-30 PMID: 1895409
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.12.7879-7890.1994
Published
1994-12-00
Pages
7879-90
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237250
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.68.12.7879-7890.1994 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2