JNK-mediated phosphorylation of paxillin in adhesion assembly and tension-induced cell death by the adenovirus death factor E4orf4.
The adenovirus type 2 Early Region 4 ORF4 (E4orf4) protein induces a caspase-independent death program in tumor cells involving changes in actin dynamics that are functionally linked to cell killing. Because an increase in myosin II-based contractility is needed for the death of E4orf4-expressing cells, we have proposed that alteration of cytoskeletal tension is part of the signals engaging the death pathway. Yet the mechanisms involved are poorly defined. Herein, we show that the Jun N-terminal kinase JNK is activated in part through a pathway involving Src, Rho, and ROCK (Rho kinase) and contributes to dysregulate adhesion dynamics and to kill cells in response to E4orf4. JNK supports the formation of atypically robust focal adhesions, which are bound to the assembly of the peculiar actomyosin network typifying E4orf4-induced cell death and which are required for driving nuclear condensation. Remarkably, the dramatic enlargement of focal adhesions, actin remodeling, and cell death all rely on paxillin phosphorylation at Ser-178, which is induced by E4orf4 in a JNK-dependent way. Furthermore, we found that Ser-178-paxillin phosphorylation is necessary to decrease adhesion turnover and to enhance the time residency of paxillin at focal adhesions, promoting its recruitment from an internal pool. Our results indicate that perturbation of tensional homeostasis by E4orf4 involves JNK-regulated changes in paxillin adhesion dynamics that are required to engage the death pathway. Moreover, our findings support a role for JNK-mediated paxillin phosphorylation in adhesion growth and stabilization during tension signaling.
Smadja-Lamère
Nicolas
N
Centre de Recherche en Cancérologie de l'Université Laval, L'Hôtel-Dieu de Québec, CRCHUQ, Québec, G1R 2J6, Canada.
Boulanger
Marie-Chloé
MC
Champagne
Claudia
C
Branton
Philip E
PE
Lavoie
Josée N
JN
D000199:Actins / Q000378:metabolism; D000256:Adenoviridae / Q000378:metabolism; D002448:Cell Adhesion; D016923:Cell Death; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D005822:Genetic Vectors; D006706:Homeostasis; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D008954:Models, Biological; D051419:Paxillin / Q000737:chemistry*; D010766:Phosphorylation; D014764:Viral Proteins / Q000737:chemistry*; D019061:src-Family Kinases / Q000378:metabolism
D000199:Actins; C121517:E4orf4 protein, adenovirus; D051419:Paxillin; D014764:Viral Proteins; D019061:src-Family Kinases; D048670:MAP Kinase Kinase 4