Jurkat control vs J Tat72

Triplicate

Jurkat control vs J Tat101

Triplicate

Jurkat Control

Jurkat Tat72

Jurkat Tat101

0.000

2.000

4.000

6.000

8.000

hsa-miR-21

Jurkat Control

Jurkat Tat72

Jurkat Tat101

0.0001.0002.0003.0004.0005.0006.000

hsa-miR-222

Jurkat Control

Jurkat Tat72

Jurkat Tat101

0.0000.2000.4000.6000.8001.0001.200

hsa-miR-128

HIV-1 Tat protein up-regulates human cellular miRNAs involved in T cell apoptosis and cell cycle regulation: requirement of Tat second exon

INTRODUCTION RESULTS

microRNA array

Confirmation by qRT-PCR

Fo

ld c

ha

ng

e

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

Jurkat Con-trol

Jurkat Tat72 Jurkat Tat101

0.0002.0004.0006.0008.000

10.00012.00014.000

premiR-21

0

10

20

30

40

50

60

70

80

90

0h 2h 4h 6h 18h

JJTetOff

JJTat72

JJTat101% A

nn

exin

-V

Bcl-2FasL 4h

J C

ontr

olJ

Tat7

2J

Tat1

01

-actin

- - - + + +

J C

ontr

olJ

Tat7

2J

Tat1

01

Jurkat Con-trol

Jurkat Tat72

Jurkat Tat101

0.0000.2000.4000.6000.8001.0001.200

PTEN

Jurkat Con-trol

Jurkat Tat72

Jurkat Tat101

0.0000.2000.4000.6000.8001.0001.200

PDCD4

Up-regulation of cellular miR-21 by Tat and its implication in apoptosis

The precursor of miR-21 is also increased

Fo

ld c

ha

ng

e

miR-21 targets are down-regulated

Fo

ld c

ha

ng

e

These proteins have pro-apoptotic effects

Increased resistance to apoptosis in Jurkat Tat101 High levels of miR-21 correlates to the increase of the

anti-apoptotic protein Bcl-2

Jurkat Con-trol

Jurkat Tat72

Jurkat Tat101

0.000

2.000

4.000

6.000

8.000

premiR-222

Jurkat Control

Jurkat Tat72

Jurkat Tat101

0.0000.2000.4000.6000.8001.0001.200

CDKN1B

Up-regulation of cellular miR-222 by Tat and its relation to cell cycle progression

The precursor of miR-222 is also increasedF

old

ch

an

ge

miR-222 target is down-regulated

CDKN1B inhibits cell

cycle progression at

G1 phase

Fo

ld c

ha

ng

e

Acknowledgements

G1/S phases: 20 % G1/S phases: 21,45 %

G1/S phases: 53,60 %

Jurkat controlPropidium iodide

Ev

en

ts

Jurkat Tat72Propidium iodide

Jurkat Tat101Propidium 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.