Abstract
Background: HIV-1 TAT protein is essential for the regulation of viral genome transcription. The first exon of TAT protein has a fundamental role in the stimulation of the extrinsic and intrinsic apoptosis pathways, but its anti-HIV activity is not clear yet.
Methods: In the current study, we firstly cloned the first exon of the TAT coding sequence in the pET-24a expression vector and then protein expression was done in the Rosetta expression host. Next, the expressed TAT protein was purified by Ni-NTA column under native conditions. After that, the protein yield was determined by Bradford kit and NanoDrop spectrophotometry. Finally, the cytotoxicity effect and anti-Scr-HIV-1 activity of the recombinant TAT protein alone and along with Tenofovir drug were assessed by MTT and ELISA, respectively.
Results: The recombinant TAT protein was successfully generated in E. coli, as confirmed by 13.5% SDS-PAGE and western blotting. The protein yield was ~150-200 μg/ml. In addition, the recombinant TAT protein at a certain dose with low toxicity could suppress Scr-HIV replication in the infected HeLa cells (~30%) that was comparable with a low toxic dose of Tenofovir drug (~40%). It was interesting that the recombinant TAT protein could enhance anti-HIV potency of Tenofovir drug up to 66%.
Conclusion: Generally, a combination of TAT protein and Tenofovir drug could significantly inhibit HIV-1 replication. It will be required to determine their mechanism of action in the next studies.
Keywords: HIV-1, TAT protein, prokaryotic expression system, highly active antiretroviral therapy, tenofovir drug, anti-HIV activity.
Graphical Abstract
[http://dx.doi.org/10.3390/v4113020] [PMID: 23202514]
[http://dx.doi.org/10.1159/000445852] [PMID: 27403093]
[http://dx.doi.org/10.1590/S0034-89102006000200023] [PMID: 16583048]
[http://dx.doi.org/10.1016/j.vaccine.2020.04.015] [PMID: 32387010]
[http://dx.doi.org/10.1128/MMBR.00065-15] [PMID: 27357278]
[http://dx.doi.org/10.1016/S0014-5793(03)00725-7] [PMID: 12885404]
[http://dx.doi.org/10.1002/prot.25609] [PMID: 30260061]
[http://dx.doi.org/10.1128/JVI.71.7.5528-5539.1997] [PMID: 9188627]
[http://dx.doi.org/10.1089/aid.2011.0230] [PMID: 21848364]
[http://dx.doi.org/10.3389/fmicb.2017.02151] [PMID: 29163435]
[http://dx.doi.org/10.1128/JVI.02128-18] [PMID: 30651369]
[http://dx.doi.org/10.1074/jbc.H117.791731] [PMID: 28986430]
[http://dx.doi.org/10.1016/j.jmb.2011.04.042] [PMID: 21762802]
[http://dx.doi.org/10.1186/1743-422X-10-358] [PMID: 24359561]
[http://dx.doi.org/10.1038/334165a0] [PMID: 3386755]
[http://dx.doi.org/10.1186/s12967-018-1500-0] [PMID: 29739413]
[http://dx.doi.org/10.1146/annurev.biochem.67.1.1] [PMID: 9759480]
[http://dx.doi.org/10.1111/j.1600-0854.2011.01286.x] [PMID: 21951552]
[http://dx.doi.org/10.1016/S0065-3527(09)74001-1] [PMID: 19698894]
[http://dx.doi.org/10.1186/s12977-017-0330-0] [PMID: 28122580]
[http://dx.doi.org/10.1007/978-3-540-33397-5_5] [PMID: 16570858]
[http://dx.doi.org/10.1073/pnas.90.7.2695] [PMID: 8464877]
[http://dx.doi.org/10.1186/1742-4690-7-114] [PMID: 21176220]
[http://dx.doi.org/10.1038/nm946] [PMID: 14528301]
[http://dx.doi.org/10.1016/S1097-2765(03)00353-8] [PMID: 14527406]
[http://dx.doi.org/10.1097/QAD.0000000000001113] [PMID: 27064995]
[http://dx.doi.org/10.1128/JVI.63.9.4110-4114.1989] [PMID: 2474678]
[http://dx.doi.org/10.3390/ijms18010126] [PMID: 28075409]
[http://dx.doi.org/10.1038/s41598-019-48689-x] [PMID: 31511615]
[http://dx.doi.org/10.3389/fmicb.2014.00232] [PMID: 24904546]
[http://dx.doi.org/10.1128/JVI.01322-19] [PMID: 31578291]
[http://dx.doi.org/10.1182/blood.V96.9.2951] [PMID: 11049971]
[http://dx.doi.org/10.1038/nature09131] [PMID: 20535204]
[http://dx.doi.org/10.2174/1381612823666170704130635] [PMID: 28677507]
[http://dx.doi.org/10.1016/j.yjmcc.2011.07.017] [PMID: 21820442]
[http://dx.doi.org/10.1371/journal.pone.0077746] [PMID: 24223723]
[http://dx.doi.org/10.1186/1742-4690-2-5] [PMID: 15691386]
[http://dx.doi.org/10.1038/s41598-018-27008-w] [PMID: 29875444]
[http://dx.doi.org/10.1002/jcp.20252] [PMID: 15573381]
[http://dx.doi.org/10.4149/av_2011_01_15] [PMID: 21434701]
[http://dx.doi.org/10.2174/1567201814666171129210654] [PMID: 29189153]
[http://dx.doi.org/10.1099/vir.0.016303-0] [PMID: 19812265]
[http://dx.doi.org/10.1073/pnas.90.16.7632] [PMID: 8395050]
[http://dx.doi.org/10.1006/abbi.2000.2197] [PMID: 11368344]
[http://dx.doi.org/10.1074/jbc.M108591200] [PMID: 11827962]
[http://dx.doi.org/10.1016/j.immuni.2005.03.010] [PMID: 15894278]
[http://dx.doi.org/10.1016/j.jviromet.2009.11.021] [PMID: 19941902]
[http://dx.doi.org/10.1007/s12250-009-3068-6]
[http://dx.doi.org/10.1006/prep.1996.0076] [PMID: 8812837]
[http://dx.doi.org/10.1016/S0014-5793(02)03624-4] [PMID: 12459459]
[http://dx.doi.org/10.1016/j.jbiotec.2006.07.012] [PMID: 16959350]
[http://dx.doi.org/10.1016/j.jbiotec.2004.08.004] [PMID: 15607230]
[http://dx.doi.org/10.1039/9781849730211]
[http://dx.doi.org/10.1016/S0076-6879(00)26058-8] [PMID: 11036646]
[http://dx.doi.org/10.1074/jbc.271.25.14819] [PMID: 8662917]
[http://dx.doi.org/10.1016/S0166-3542(01)00210-8] [PMID: 11888656]
[http://dx.doi.org/10.3390/ijms18030531] [PMID: 28257038]
[http://dx.doi.org/10.1177/095632029500600403]
[http://dx.doi.org/10.1080/22221751.2020.1752609] [PMID: 32267205]
[http://dx.doi.org/10.1126/science.368.6493.807] [PMID: 32439768]
[http://dx.doi.org/10.2174/0929866526666190823145916] [PMID: 31441722]
[http://dx.doi.org/10.2174/1389201020666190729104648] [PMID: 31362669]
[http://dx.doi.org/10.1038/srep41018] [PMID: 28145455]
[http://dx.doi.org/10.4049/jimmunol.181.12.8460] [PMID: 19050264]
[http://dx.doi.org/10.2174/1381612824666180608124549] [PMID: 29886823]
[http://dx.doi.org/10.3109/21691401.2014.883400] [PMID: 24564348]
[http://dx.doi.org/10.1016/j.ijpharm.2017.02.038] [PMID: 28216464]
[http://dx.doi.org/10.1002/prp2.188] [PMID: 26516592]
[http://dx.doi.org/10.18433/J36W3X] [PMID: 26517135]
[http://dx.doi.org/10.1016/j.jconrel.2014.07.049] [PMID: 25094032]
[http://dx.doi.org/10.1016/j.jconrel.2011.11.021] [PMID: 22138068]
[http://dx.doi.org/10.1016/j.addr.2016.02.006] [PMID: 26928656]
[http://dx.doi.org/10.1016/j.jddst.2019.03.009]