The localization of prenylated Ras at the plasma membrane promotes activation of Ras by receptor tyrosine kinases and stimulates oncogenic signaling by mutant Ras. The Nogo-B receptor (NgBR) is a transmembrane receptor that contains a conserved hydrophobic pocket. Here, we demonstrate that the NgBR promotes the membrane accumulation of Ras by directly binding prenylated Ras at the plasma membrane. We show that NgBR knockdown diminishes the membrane localization of Ras in multiple cell types. NgBR overexpression in NIH-3T3 fibroblasts increases membrane-associated Ras, induces the transformed phenotype in vitro, and promotes the formation of fibrosarcoma in nude mice. NgBR knockdown in human breast cancer cells reduces Ras membrane localization, inhibits epidermal growth factor (EGF)-stimulated Ras signaling and diminishes tumorigenesis of xenografts in nude mice. Our data demonstrate that NgBR is a unique receptor that promotes accumulation of prenylated Ras at the plasma membrane and promotes EGF pathways.
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Dr John Hancock at University of Texas Health Science Center at Houston generously provided pEGFP-C1 vector. This work is supported in part by start-up funds from Division of Pediatric Surgery and Division of Pediatric Pathology, Medical College of Wisconsin (MCW) and Advancing a Healthier Wisconsin endowment to MCW, AHA SDG 0730079N, NIH R01HL108938, Wisconsin Breast Cancer Showhouse (WBCS), Institutional Research Grant # 86-004-26 from the American Cancer Society, We Care Fund, Kathy Duffey Fogarty Award for breast cancer research, State of Wisconsin Tax Check-off program for breast & prostate cancer research and Children’s Hospital of Wisconsin Research Institute Pilot Innovative Research Grant to QRM., AHA postdoctoral fellowship 13POST13940002 and 11POST5690035, China State Key Basic Research Program Grant 2016YFA0501401 and the support from the Hundred Talents Program of CAS to BZ. Additional support was provided by NIH R01 CA188871 (CLW), the Rock River Cancer Research Foundation (CLW), and the Nancy Laning Sobczak, PhD., Breast Cancer Research Award (CLW), NIH R01HL128647 (ZY).
BZ, WH, SK, PG, UR, ZL, BW, WQD, QRM conducted the experiments; QRM, BZ, WH designed the experiments and wrote the paper; ZY provided computer modeling; CW provided reagents and edited the paper; QRM was responsible for overall integration and execution of the scientific approaches.
The authors declare no conflict of interest.
Supplementary Information accompanies this paper on the Oncogene website
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