TY - JOUR
T1 - Receptor-interacting Ser/Thr kinase 1 (RIPK1) and myosin IIA– dependent ceramidosomes form membrane pores that mediate blebbing and necroptosis
AU - Nganga, Rose
AU - Oleinik, Natalia
AU - Kim, Jisun
AU - Selvam, Shanmugam Panneer
AU - De Palma, Ryan
AU - Johnson, Kristen A.
AU - Parikh, Rasesh Y.
AU - Gangaraju, Vamsi
AU - Peterson, Yuri
AU - Dany, Mohammed
AU - Stahelin, Robert V.
AU - Voelkel-Johnson, Christina
AU - Szulc, Zdzislaw M.
AU - Bieberich, Erhard
AU - Ogretmen, Besim
N1 - Publisher Copyright:
© 2019 Nganga et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2019/1/11
Y1 - 2019/1/11
N2 - Formation of membrane pores/channels regulates various cellular processes, such as necroptosis or stem cell niche signaling. However, the roles of membrane lipids in the formation of pores and their biological functions are largely unknown. Here, using the cellular stress model evoked by the sphingolipid analog drug FTY720, we show that formation of ceramide-enriched membrane pores, referred to here as ceramidosomes, is initiated by a receptor-interacting Ser/Thr kinase 1 (RIPK1)– ceramide complex transported to the plasma membrane by nonmuscle myosin IIA– dependent trafficking in human lung cancer cells. Molecular modeling/simulation coupled with site-directed mutagenesis revealed that Asp147 or Asn169 of RIPK1 are key for ceramide binding and that Arg258 or Leu293 residues are involved in the myosin IIA interaction, leading to ceramidosome formation and necroptosis. Moreover, generation of ceramidosomes independently of any external drug/stress stimuli was also detected in the plasma membrane of germ line stem cells in ovaries during the early stages of oogenesis in Drosophila melanogaster. Inhibition of ceramidosome formation via myosin IIA silencing limited germ line stem cell signaling and abrogated oogenesis. In conclusion, our findings indicate that the RIPK1– ceramide complex forms large membrane pores we named ceramidosomes. They further suggest that, in addition to their roles in stress-mediated necroptosis, these ceramide-enriched pores also regulate membrane integrity and signaling and might also play a role in D. melanogaster ovary development.
AB - Formation of membrane pores/channels regulates various cellular processes, such as necroptosis or stem cell niche signaling. However, the roles of membrane lipids in the formation of pores and their biological functions are largely unknown. Here, using the cellular stress model evoked by the sphingolipid analog drug FTY720, we show that formation of ceramide-enriched membrane pores, referred to here as ceramidosomes, is initiated by a receptor-interacting Ser/Thr kinase 1 (RIPK1)– ceramide complex transported to the plasma membrane by nonmuscle myosin IIA– dependent trafficking in human lung cancer cells. Molecular modeling/simulation coupled with site-directed mutagenesis revealed that Asp147 or Asn169 of RIPK1 are key for ceramide binding and that Arg258 or Leu293 residues are involved in the myosin IIA interaction, leading to ceramidosome formation and necroptosis. Moreover, generation of ceramidosomes independently of any external drug/stress stimuli was also detected in the plasma membrane of germ line stem cells in ovaries during the early stages of oogenesis in Drosophila melanogaster. Inhibition of ceramidosome formation via myosin IIA silencing limited germ line stem cell signaling and abrogated oogenesis. In conclusion, our findings indicate that the RIPK1– ceramide complex forms large membrane pores we named ceramidosomes. They further suggest that, in addition to their roles in stress-mediated necroptosis, these ceramide-enriched pores also regulate membrane integrity and signaling and might also play a role in D. melanogaster ovary development.
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U2 - 10.1074/jbc.RA118.005865
DO - 10.1074/jbc.RA118.005865
M3 - Article
C2 - 30420430
AN - SCOPUS:85059844392
SN - 0021-9258
VL - 294
SP - 502
EP - 519
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 2
ER -