DOI: 10.1128/jvi.73.11.9011-9020.1999. Structure-Based Mutagenesis of the Human Immunodeficiency Virus Type 1 DNA Attachment Site: Effects on Integration and cDNA Synthesis

Structure-Based Mutagenesis of the Human Immunodeficiency Virus Type 1 DNA Attachment Site: Effects on Integration and cDNA Synthesis

10.1128/jvi.73.11.9011-9020.1999

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

Abstract

ABSTRACTSequences at the ends of linear retroviral cDNA important for integration define the viral DNA attachment (att) site. Whereas determinants of human immunodeficiency virus type 1 (HIV-1) integrase important for replication in T lymphocytes have been extensively characterized, regions of theattsite important for viral spread have not been thoroughly examined. Previous transposon-mediated footprinting of preintegration complexes isolated from infected cells revealed enhanced regions of bacteriophage Mu insertion near the ends of HIV-1 cDNA, in the regions of theattsites. Here, we identified the subterminal cDNA sequences cleaved during in vitro footprinting and used this structure-based information together with results of previous work to construct and characterize 24attsite mutant viruses. We found that although subterminal cDNA sequences contributed to HIV-1 replication, the identities of these bases were not critical for integration. In contrast, the phylogenetically conserved CA dinucleotides located at the ends of HIV-1 contributed significantly to virus replication and integration. Mutants containing one intact CA end displayed delays in peak virus growth compared to the wild type. In contrast, double mutant viruses lacking both CAs were replication defective. The A of the CA appeared to be the most critical determinant of integration, because two different U5 mutant viruses containing the substitution of TG for CA partially reverted by changing the G back to A. We also identified a U5 deletion mutant in which the CA played a crucial role in reverse transcription.

Bibliography

Brown, H. E. V., Chen, H., & Engelman, A. (1999). Structure-Based Mutagenesis of the Human Immunodeficiency Virus Type 1 DNA Attachment Site: Effects on Integration and cDNA Synthesis. Journal of Virology, 73(11), 9011â9020.

Authors 3

Heidi E. V. Brown (first)
Hongmin Chen (additional)
Alan Engelman (additional)

References 38 Referenced 74

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Brown P. O. Integration Retroviruses. Coffin J. M. Hughes S. H. Varmus H. E. 1997 161 203 Cold Spring Harbor Laboratory Cold Spring Harbor N.Y
10.1016/0092-8674(87)90287-X
10.1074/jbc.274.24.17358
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10.1128/jvi.65.8.4350-4358.1991
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Sambrook J. Fritsch E. F. Maniatis T. Molecular cloning: a laboratory manual 2nd ed. 1989 Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y
Telesnitsky A. Goff S. P. Reverse transcriptase and the generation of retroviral DNA Retroviruses. Coffin J. M. Hughes S. H. Varmus H. E. 1997 121 160 Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y
10.1093/emboj/16.24.7511
10.1073/pnas.95.18.10535
Weiss A. Wiskocil R. L. Stobo J. D. The role of T3 surface molecules in the activation of human T cells: a two-stimulus requirement for IL 2 production reflects events occurring at a pretranslational level.J. Immunol. 133 1984 123 128 (10.4049/jimmunol.133.1.123) / J. Immunol. / The role of T3 surface molecules in the activation of human T cells: a two-stimulus requirement for IL 2 production reflects events occurring at a pretranslational level by Weiss A. (1984)
10.1093/nar/9.1.133

Dates

Type	When
Created	5 years, 7 months ago (Dec. 31, 2019, 1:20 p.m.)
Deposited	1 year, 11 months ago (Sept. 24, 2023, 11:08 a.m.)
Indexed	3 months, 1 week ago (May 21, 2025, 7:43 a.m.)
Issued	25 years, 9 months ago (Nov. 1, 1999)
Published	25 years, 9 months ago (Nov. 1, 1999)
Published Print	25 years, 9 months ago (Nov. 1, 1999)

Funders 0

None

BibTeX

@article{Brown_1999, title={Structure-Based Mutagenesis of the Human Immunodeficiency Virus Type 1 DNA Attachment Site: Effects on Integration and cDNA Synthesis}, volume={73}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.73.11.9011-9020.1999}, DOI={10.1128/jvi.73.11.9011-9020.1999}, number={11}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Brown, Heidi E. V. and Chen, Hongmin and Engelman, Alan}, year={1999}, month=nov, pages={9011–9020} }

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