HAL Id: inserm-00826959
https://www.hal.inserm.fr/inserm-00826959
Submitted on 16 May 2014
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To cite this version:
Sylvie Elsen, Véronique Collin-Faure, Xavier Gidrol, Claudie Lemercier. The opportunistic pathogen Pseudomonas aeruginosa activates the DNA double-strand break signaling and repair pathway in infected cells.. Cellular and Molecular Life Sciences, Springer Verlag, 2013, 70 (22), pp.4385-97. �10.1007/s00018-013-1392-3�. �inserm-00826959�
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c-Jun S63 m o c k Pa mo c k Pa mo c k Pa c-Jun no CIP + CIP + CIP + inhib γγγγH2AX GAPDH C M C M Pa c-Jun S63 c-Jun GAPDH c-Jun S73 mock Relative c-Jun intensity 0 1 2 3a
b
e
C M C M C M C M 0 1 5 10 MOI c-Jun γγγγH2AX GAPDH 0 1 2 3 Relative γγγγH2AX intensity C M C M C M m o c k C H A PA O 1 GAPDH C M C M PA K E. c o li c-Jun γγγγH2AX 0 2 4 6 8 10 Relative γγγγH2AX intensityd
0 10 20 30 40 50 60 Eto % o f a p o p to ti c o r d e a d c e ll s dead/necrosis apoptotic 1h 2h30 5h Time CHA - - CHA - CHAc
γγγγ H 2 A X l e v e l (f o ld in c re a s e ) mock Pa 0 1 5 10 15 C M C M C M C M 1h γγγγH2AX mock MEF2D GAPDH 2h30 5h c-Jun Pa Relative γγγγH2AX intensity 0 20 40 120mock
53 BP1
γγγγ
H2AX
53 BP1 +
γγγγ
H2AX
DNA
MOI
1
MOI
10
MOI
100
Irrad.
2 Gy
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*
*
*
*
*
*
*
0 10 20 30 40 50 60 Mock Mock H2O2 MOI 10 MOI 100 % o f D N A i n t a il Mock Mock H2O2 MOI 100 MOI 10
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Mock Mock H2O2 MOI 10 MOI 100 % o f c e ll s w it h c o m e t ta il 0 20 40 60 80 100ExoS C H A S SRK S2ED CHA ∆∆∆∆ExoS
-b
d
e
a
c
ADP-ribosyltransferase Rho GAP Sec Chap 146 379 381 RK 2ED C M C M C Mmock CHA ∆∆∆∆CHAExsA
0 2 4 6 8 10 12 c-Jun GAPDH γγγγH2AX Relative γγγγH2AX intensity CHA mock WT ∆∆∆∆S ∆∆∆∆T ∆∆∆∆ST c-Jun γγγγH2AX GAPDH 6 8 0 2 4 Relative γγγγH2AX intensity m o c k C H A S S 2ED S RK c-Jun γγγγH2AX GAPDH CHA ∆∆∆∆ExoS -0.0 0.2 0.4 0.6 0.8 1.0 1.2 Relative γγγγH2Ax intensity
GAPDH m o c k γγγγH2AX C H A C H A ∆∆∆∆ S m o c k C H A C H A∆∆∆∆ S m o c k C H A C H A ∆∆∆∆ S 1h 2h30 5h pATM ATM 53BP1
a
b
c
d
m o c k GAPDH C H A C H A ∆∆∆∆ S m o c k C H A C H A ∆∆∆∆ S pATM + ATM inh Inhibitor γγγγH2AX GAPDH c-Jun J N K i n h P3 8 i n h Er k i n h A T M i n h D M SO J N K i n h P3 8 i n h Er k i n h A T M i n h D M SO mock Pa Relative γγγγH2AX intensity 0.0 0.2 0.4 0.6 0.8 1.0 1.2 pATM 53BP1 ATM inh - + - + 0 Gy 2 Gy ATM γγγγH2AX GAPDH 0 1 2 3 4 Relative pATM intensityb
53 BP1
γγγγH2AX 53 BP1 + γγγγH2AX DNA
mock CHA 2 Gy
