In vitro Anti-HIV-1 Activity of the Recombinant HIV-1 TAT Protein Along With Tenofovir Drug

Author(s): Maryam-Sadat Yadavar-Nikravesh, Alireza Milani, Rouhollah Vahabpour, Mehdi Khoobi, Haleh Bakhshandeh*, Azam Bolhassani*

Journal Name: Current HIV Research

Volume 19 , Issue 2 , 2021


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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.

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Article Details

VOLUME: 19
ISSUE: 2
Year: 2021
Published on: 26 February, 2021
Page: [138 - 146]
Pages: 9
DOI: 10.2174/1570162X18666201012152600
Price: $65

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