1. Jurkat control vs J Tat72 Triplicate Jurkat control vs J Tat101 Triplicate HIV-1 Tat protein up-regulates human cellular miRNAs involved in T cell apoptosis and cell cycle regulation: requirement of Tat second exon INTRODUCTIONRESULTS microRNA array Confirmation by qRT-PCR Fold change We observe that… 1- HIV-1 Tat does not show RNAi suppressor activity 2- Tat specifically dysregulates some cellular miRNAs 3- These changes are dependent on the second exon of Tat

2. % Annexin-V Bcl-2 FasL 4h J ControlJ Tat72J Tat101  -actin - - - + + + J ControlJ Tat72J Tat101 Up-regulation of cellular miR-21 by Tat and its implication in apoptosis The precursor of miR-21 is also increased Fold change miR-21 targets are down-regulated Fold change These proteins have pro-apoptotic effects Increased resistance to apoptosis in Jurkat Tat101High levels of miR-21 correlates to the increase of the anti-apoptotic protein Bcl-2

3. Up-regulation of cellular miR-222 by Tat and its relation to cell cycle progression The precursor of miR-222 is also increased Fold change miR-222 target is down-regulated CDKN1B inhibits cell cycle progression at G1 phase Fold change Acknowledgements G1/S phases: 20 % G1/S phases: 21,45 % G1/S phases: 53,60 % Jurkat control Propidium iodide Events Jurkat Tat72 Propidium iodide Jurkat Tat101 Propidium iodide Increased activation of cell cycle in Jurkat Tat101 at G1 and early S phases Conclusions -HIV-1 Tat does not show RNAi suppressor activity, but it specifically up-regulates miR-21 and miR-222 and their precursors and down- regulates miR-128 -These modifications are dependent on the second exon of Tat. -Targets of the enhanced miRNAs are down-regulated. This could explain the specific phenotype of resistance to apoptosis and cell cycle activation observed in Jurkat Tat101.