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TRAF2 recruitment via T61 in CD30 drives NF?B activation and enhances hESC survival and proliferation.

ABSTRACT: CD30 (TNFRSF8), a tumor necrosis factor receptor family protein, and CD30 variant (CD30v), a ligand-independent form encoding only the cytoplasmic signaling domain, are concurrently overexpressed in transformed human embryonic stem cells (hESCs) or hESCs cultured in the presence of ascorbate. CD30 and CD30v are believed to increase hESC survival and proliferation through NF?B activation, but how this occurs is largely unknown. Here we demonstrate that hESCs that endogenously express CD30v and hESCs that artificially overexpress CD30v exhibit increased ERK phosphorylation levels, activation of the canonical NF?B pathway, down-regulation of the noncanonical NF?B pathway, and reduced expression of the full-length CD30 protein. We further find that CD30v, surprisingly, resides predominantly in the nucleus of hESC. We demonstrate that alanine substitution of a single threonine residue at position 61 (T61) in CD30v abrogates CD30v-mediated NF?B activation, CD30v-mediated resistance to apoptosis, and CD30v-enhanced proliferation, as well as restores normal G2/M-checkpoint arrest upon H2O2 treatment while maintaining its unexpected subcellular distribution. Using an affinity purification strategy and LC-MS, we identified TRAF2 as the predominant protein that interacts with WT CD30v but not the T61A-mutant form in hESCs. The identification of Thr-61 as a critical residue for TRAF2 recruitment and canonical NF?B signaling by CD30v reveals the substantial contribution that this molecule makes to overall NF?B activity, cell cycle changes, and survival in hESCs.

SUBMITTER: Thakar NY

PROVIDER: S-EPMC4342033 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature

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TRAF2 recruitment via T61 in CD30 drives NFκB activation and enhances hESC survival and proliferation.

Thakar Nilay Y NY Ovchinnikov Dmitry A DA Hastie Marcus L ML Kobe Bostjan B Gorman Jeffrey J JJ Wolvetang Ernst J EJ

Molecular biology of the cell 20150107 5

CD30 (TNFRSF8), a tumor necrosis factor receptor family protein, and CD30 variant (CD30v), a ligand-independent form encoding only the cytoplasmic signaling domain, are concurrently overexpressed in transformed human embryonic stem cells (hESCs) or hESCs cultured in the presence of ascorbate. CD30 and CD30v are believed to increase hESC survival and proliferation through NFκB activation, but how this occurs is largely unknown. Here we demonstrate that hESCs that endogenously express CD30v and hE ...[more]

PMID: 25568342

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Dataset Information

TRAF2 recruitment via T61 in CD30 drives NF?B activation and enhances hESC survival and proliferation.

Publications

TRAF2 recruitment via T61 in CD30 drives NFκB activation and enhances hESC survival and proliferation.

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