| Structural highlights
Function
[POL_HV1H2] Gag-Pol polyprotein and Gag polyprotein may regulate their own translation, by the binding genomic RNA in the 5'-UTR. At low concentration, Gag-Pol and Gag would promote translation, whereas at high concentration, the polyproteins encapsidate genomic RNA and then shutt off translation.[1] [2] [3] [4] Matrix protein p17 has two main functions: in infected cell, it targets Gag and Gag-pol polyproteins to the plasma membrane via a multipartite membrane-binding signal, that includes its myristoylated N-terminus. The second function is to play a role in nuclear localization of the viral genome at the very start of cell infection. Matrix protein is the part of the pre-integration complex. It binds in the cytoplasm the human BAF protein which prevent autointegration of the viral genome, and might be included in virions at the ration of zero to 3 BAF dimer per virion. The myristoylation signal and the NLS thus exert conflicting influences its subcellular localization. The key regulation of these motifs might be phosphorylation of a portion of MA molecules on the C-terminal tyrosine at the time of virus maturation, by virion-associated cellular tyrosine kinase. Implicated in the release from host cell mediated by Vpu.[5] [6] [7] [8] Capsid protein p24 forms the conical core that encapsulates the genomic RNA-nucleocapsid complex in the virion. Most core are conical, with only 7% tubular. The core is constituted by capsid protein hexamer subunits. The core is disassembled soon after virion entry. Interaction with human PPIA/CYPA protects the virus from restriction by human TRIM5-alpha and from an unknown antiviral activity in human cells. This capsid restriction by TRIM5 is one of the factors which restricts HIV-1 to the human species.[9] [10] [11] [12] Nucleocapsid protein p7 encapsulates and protects viral dimeric unspliced (genomic) RNA. Binds these RNAs through its zinc fingers. Facilitates rearangement of nucleic acid secondary structure during retrotranscription of genomic RNA. This capability is referred to as nucleic acid chaperone activity.[13] [14] [15] [16] The aspartyl protease mediates proteolytic cleavages of Gag and Gag-Pol polyproteins during or shortly after the release of the virion from the plasma membrane. Cleavages take place as an ordered, step-wise cascade to yield mature proteins. This process is called maturation. Displays maximal activity during the budding process just prior to particle release from the cell. Also cleaves Nef and Vif, probably concomitantly with viral structural proteins on maturation of virus particles (By similarity).[17] [18] [19] [20] Reverse transcriptase/ribonuclease H (RT) is a multifunctional enzyme that converts the viral RNA genome into dsDNA in the cytoplasm, shortly after virus entry into the cell. This enzyme displays a DNA polymerase activity that can copy either DNA or RNA templates, and a ribonuclease H (RNase H) activity that cleaves the RNA strand of RNA-DNA heteroduplexes in a partially processive 3' to 5' endonucleasic mode. Conversion of viral genomic RNA into dsDNA requires many steps. A tRNA(3)-Lys binds to the primer-binding site (PBS) situated at the 5'-end of the viral RNA. RT uses the 3' end of the tRNA primer to perform a short round of RNA-dependent minus-strand DNA synthesis. The reading proceeds through the U5 region and ends after the repeated (R) region which is present at both ends of viral RNA. The portion of the RNA-DNA heteroduplex is digested by the RNase H, resulting in a ssDNA product attached to the tRNA primer. This ssDNA/tRNA hybridizes with the identical R region situated at the 3' end of viral RNA. This template exchange, known as minus-strand DNA strong stop transfer, can be either intra- or intermolecular. RT uses the 3' end of this newly synthesized short ssDNA to perform the RNA-dependent minus-strand DNA synthesis of the whole template. RNase H digests the RNA template except for two polypurine tracts (PPTs) situated at the 5'-end and near the center of the genome. It is not clear if both polymerase and RNase H activities are simultaneous. RNase H probably can proceed both in a polymerase-dependent (RNA cut into small fragments by the same RT performing DNA synthesis) and a polymerase-independent mode (cleavage of remaining RNA fragments by free RTs). Secondly, RT performs DNA-directed plus-strand DNA synthesis using the PPTs that have not been removed by RNase H as primers. PPTs and tRNA primers are then removed by RNase H. The 3' and 5' ssDNA PBS regions hybridize to form a circular dsDNA intermediate. Strand displacement synthesis by RT to the PBS and PPT ends produces a blunt ended, linear dsDNA copy of the viral genome that includes long terminal repeats (LTRs) at both ends.[21] [22] [23] [24] Integrase catalyzes viral DNA integration into the host chromosome, by performing a series of DNA cutting and joining reactions. This enzyme activity takes place after virion entry into a cell and reverse transcription of the RNA genome in dsDNA. The first step in the integration process is 3' processing. This step requires a complex comprising the viral genome, matrix protein, Vpr and integrase. This complex is called the pre-integration complex (PIC). The integrase protein removes 2 nucleotides from each 3' end of the viral DNA, leaving recessed CA OH's at the 3' ends. In the second step, the PIC enters cell nucleus. This process is mediated through integrase and Vpr proteins, and allows the virus to infect a non dividing cell. This ability to enter the nucleus is specific of lentiviruses, other retroviruses cannot and rely on cell division to access cell chromosomes. In the third step, termed strand transfer, the integrase protein joins the previously processed 3' ends to the 5' ends of strands of target cellular DNA at the site of integration. The 5'-ends are produced by integrase-catalyzed staggered cuts, 5 bp apart. A Y-shaped, gapped, recombination intermediate results, with the 5'-ends of the viral DNA strands and the 3' ends of target DNA strands remaining unjoined, flanking a gap of 5 bp. The last step is viral DNA integration into host chromosome. This involves host DNA repair synthesis in which the 5 bp gaps between the unjoined strands are filled in and then ligated. Since this process occurs at both cuts flanking the HIV genome, a 5 bp duplication of host DNA is produced at the ends of HIV-1 integration. Alternatively, Integrase may catalyze the excision of viral DNA just after strand transfer, this is termed disintegration.[25] [26] [27] [28]
Publication Abstract from PubMed
The retrovirus HIV-1 has been a major health issue since its discovery in the early 80s. In 2017, over 37 million people were infected with HIV-1, of which 1.8 million were new infections that year. Currently, the most successful treatment regimen is the highly active anti-retroviral therapy (HAART), which consists of a combination of three to four of the current 26 FDA-approved HIV-1 drugs. Half of these drugs target the reverse transcriptase (RT) enzyme that is essential for viral replication. One class of RT inhibitors are nucleoside reverse transcriptase inhibitors (NRTIs), a crucial component of the HAART therapy. Once incorporated into DNA, NRTIs function as a chain terminator to stop viral DNA replication. Unfortunately, treatment with NRTIs is sometimes linked to toxicity caused by off-target side effects. NRTIs may also target the replicative human mitochondrial DNA polymerase (Pol gamma), causing long-term severe drug toxicity. The goal of this work is to understand the discrimination mechanism of different NRTI analogs by RT. Crystal structures and kinetic experiments are essential for the rational design of new molecules that are able to bind selectively to RT and not Pol gamma. Structural comparison of NRTI-binding modes with both RT and Pol gamma enzymes highlights key amino acids that are responsible for the difference in affinity of these drugs to their targets. Therefore, the long-term goal of this research is to develop safer, next generation therapeutics that can overcome off-target toxicity. This article is protected by copyright. All rights reserved.
Structural insights into the recognition of Nucleoside Reverse Transcriptase Inhibitors (NRTIs) by HIV-1 reverse transcriptase: First crystal structures with RT and the active triphosphate forms of lamivudine and emtricitabine.,Bertoletti N, Chan AH, Schinazi RF, Yin YW, Anderson KS Protein Sci. 2019 Jul 13. doi: 10.1002/pro.3681. PMID:31301259[29]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Braaten D, Franke EK, Luban J. Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol. 1996 Jun;70(6):3551-60. PMID:8648689
- ↑ Anderson EC, Lever AM. Human immunodeficiency virus type 1 Gag polyprotein modulates its own translation. J Virol. 2006 Nov;80(21):10478-86. PMID:17041220 doi:80/21/10478
- ↑ Hagan NA, Fabris D. Dissecting the protein-RNA and RNA-RNA interactions in the nucleocapsid-mediated dimerization and isomerization of HIV-1 stemloop 1. J Mol Biol. 2007 Jan 12;365(2):396-410. Epub 2006 Oct 3. PMID:17070549 doi:http://dx.doi.org/10.1016/j.jmb.2006.09.081
- ↑ Ao Z, Danappa Jayappa K, Wang B, Zheng Y, Kung S, Rassart E, Depping R, Kohler M, Cohen EA, Yao X. Importin alpha3 interacts with HIV-1 integrase and contributes to HIV-1 nuclear import and replication. J Virol. 2010 Sep;84(17):8650-63. doi: 10.1128/JVI.00508-10. Epub 2010 Jun 16. PMID:20554775 doi:10.1128/JVI.00508-10
- ↑ Bertoletti N, Chan AH, Schinazi RF, Yin YW, Anderson KS. Structural insights into the recognition of Nucleoside Reverse Transcriptase Inhibitors (NRTIs) by HIV-1 reverse transcriptase: First crystal structures with RT and the active triphosphate forms of lamivudine and emtricitabine. Protein Sci. 2019 Jul 13. doi: 10.1002/pro.3681. PMID:31301259 doi:http://dx.doi.org/10.1002/pro.3681
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