TY - JOUR

T1 - Delineating the role of c-FLIP/NEMO interaction in the CD95 network via rational design of molecular probes

AU - Ivanisenko, Nikita V.

AU - Buchbinder, Jörn H.

AU - Espe, Johannes

AU - Richter, Max

AU - Bollmann, Miriam

AU - Hillert, Laura K.

AU - Ivanisenko, Vladimir A.

AU - Lavrik, Inna N.

PY - 2019/5/8

Y1 - 2019/5/8

N2 - Background: Structural homology modeling supported by bioinformatics analysis plays a key role in uncovering new molecular interactions within gene regulatory networks. Here, we have applied this powerful approach to analyze the molecular interactions orchestrating death receptor signaling networks. In particular, we focused on the molecular mechanisms of CD95-mediated NF-κB activation and the role of c-FLIP/NEMO interaction in the induction of this pathway. Results: To this end, we have created the homology model of the c-FLIP/NEMO complex using the reported structure of the v-FLIP/NEMO complex, and rationally designed peptides targeting this complex. The designed peptides were based on the NEMO structure. Strikingly, the experimental in vitro validation demonstrated that the best inhibitory effects on CD95-mediated NF-κB activation are exhibited by the NEMO-derived peptides with the substitution D242Y of NEMO. Furthermore, we have assumed that the c-FLIP/NEMO complex is recruited to the DED filaments formed upon CD95 activation and validated this assumption in silico. Further insight into the function of c-FLIP/NEMO complex was provided by the analysis of evolutionary conservation of interacting regions which demonstrated that this interaction is common in distinct mammalian species. Conclusions: Taken together, using a combination of bioinformatics and experimental approaches we obtained new insights into CD95-mediated NF-κB activation, providing manifold possibilities for targeting the death receptor network.

AB - Background: Structural homology modeling supported by bioinformatics analysis plays a key role in uncovering new molecular interactions within gene regulatory networks. Here, we have applied this powerful approach to analyze the molecular interactions orchestrating death receptor signaling networks. In particular, we focused on the molecular mechanisms of CD95-mediated NF-κB activation and the role of c-FLIP/NEMO interaction in the induction of this pathway. Results: To this end, we have created the homology model of the c-FLIP/NEMO complex using the reported structure of the v-FLIP/NEMO complex, and rationally designed peptides targeting this complex. The designed peptides were based on the NEMO structure. Strikingly, the experimental in vitro validation demonstrated that the best inhibitory effects on CD95-mediated NF-κB activation are exhibited by the NEMO-derived peptides with the substitution D242Y of NEMO. Furthermore, we have assumed that the c-FLIP/NEMO complex is recruited to the DED filaments formed upon CD95 activation and validated this assumption in silico. Further insight into the function of c-FLIP/NEMO complex was provided by the analysis of evolutionary conservation of interacting regions which demonstrated that this interaction is common in distinct mammalian species. Conclusions: Taken together, using a combination of bioinformatics and experimental approaches we obtained new insights into CD95-mediated NF-κB activation, providing manifold possibilities for targeting the death receptor network.

KW - C-FLIP

KW - Death receptor network

KW - Evolutionary conservation

KW - Homology modeling

KW - In silico

KW - NEMO

KW - NF-κB

KW - V-FLIP

KW - GAMMA

KW - APOPTOSIS

KW - KAPPA-B ACTIVATION

KW - COMPLEX

KW - PROTEIN

KW - NF-B

KW - DEATH

KW - MECHANISMS

KW - KINASE IKK

KW - REVEALS

UR - http://www.scopus.com/inward/record.url?scp=85065428080&partnerID=8YFLogxK

U2 - 10.1186/s12864-019-5539-y

DO - 10.1186/s12864-019-5539-y

M3 - Article

C2 - 31815628

AN - SCOPUS:85065428080

VL - 20

JO - BMC Genomics

JF - BMC Genomics

SN - 1471-2164

IS - Suppl 3

M1 - 293

ER -