Jurkat control vs J Tat72
description
Transcript of Jurkat control vs J Tat72
Jurkat control vs J Tat72
Triplicate
Jurkat control vs J Tat101
Triplicate
Jurkat Control
Jurkat Tat72
Jurkat Tat101
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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 RESULTSmicroRNA array
Confirmation by qRT-PCR
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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
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JJTetOffJJTat72JJTat101%
Ann
exin
-V
Bcl-2FasL 4h
J C
ontro
lJ
Tat7
2J
Tat1
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-actin
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J C
ontro
lJ
Tat7
2J
Tat1
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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
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miR-21 targets are down-regulated
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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
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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 increasedFo
ld c
hang
emiR-222 target is down-regulated
CDKN1B inhibits cell
cycle progression at
G1 phase
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Acknowledgements
G1/S phases: 20 % G1/S phases: 21,45 %
G1/S phases: 53,60 %
Jurkat controlPropidium iodide
Even
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.