RANK receptor oligomerisation in the regulation of NFκB signalling

S Das, I Sepahi, A Duthie, S Clark, J C Crockett

Research output: Contribution to journalArticle

2 Citations (Scopus)
4 Downloads (Pure)

Abstract

The interaction of receptor activator of NFκB (RANK), a member of the tumour necrosis factor receptor superfamily, with RANK ligand is crucial for the formation, function and survival of osteoclasts. The role of the cytoplasmic oligomerisation domain (pre-ligand assembly domain; PLAD or 'IVVY' motif) in the ligand-dependent activation of downstream NFκB signalling has not been studied previously. The discovery of truncating mutations of TNFRSF11A (W434X and G280X that lack the PLAD) as the cause of rare cases of osteoclast-poor osteopetrosis offered the opportunity for functional study of this region. Recapitulating the W434X mutation by transcription activator-like effector nuclease (TALEN)-mediated targeted disruption of Tnfrsf11a within the region homologous to W434X in the mouse macrophage-like cell line RAW264.7 impaired formation of osteoclast-like cells. Using overexpression studies, we demonstrated that, in contrast to WT-RANK, the absence of the PLAD in G280X-RANK and W434X-RANK prevented ligand-independent but not ligand-dependent oligomerisation. Cells expressing W434X-RANK, in which only two of the three TRAF6-binding motifs are present, continued to exhibit ligand-dependent NFκB signalling. Hence, the absence of the PLAD did not prevent ligand-induced trimerisation and subsequent NFκB activation of RANK, demonstrating that therapeutic targeting of the PLAD in the prevention of osteoporosis may not be as effective as proposed previously.

Original languageEnglish
Pages (from-to)81-91
Number of pages11
JournalJournal of Molecular Endocrinology
Volume53
Issue number1
Early online date23 May 2014
DOIs
Publication statusPublished - 1 Aug 2014

Fingerprint

Ligands
RANK Ligand
Osteoclasts
TNF Receptor-Associated Factor 6
Mutation
Tumor Necrosis Factor Receptors
Osteoporosis
Macrophages
Cell Line
Therapeutics

Keywords

  • oligomerisation
  • RANK
  • NFκB
  • osteoperosis

Cite this

RANK receptor oligomerisation in the regulation of NFκB signalling. / Das, S; Sepahi, I; Duthie, A; Clark, S; Crockett, J C.

In: Journal of Molecular Endocrinology, Vol. 53, No. 1, 01.08.2014, p. 81-91.

Research output: Contribution to journalArticle

Das, S, Sepahi, I, Duthie, A, Clark, S & Crockett, JC 2014, 'RANK receptor oligomerisation in the regulation of NFκB signalling', Journal of Molecular Endocrinology, vol. 53, no. 1, pp. 81-91. https://doi.org/10.1530/JME-14-0077
Das, S ; Sepahi, I ; Duthie, A ; Clark, S ; Crockett, J C. / RANK receptor oligomerisation in the regulation of NFκB signalling. In: Journal of Molecular Endocrinology. 2014 ; Vol. 53, No. 1. pp. 81-91.
@article{ed507f7597214d3483ec11c81d328e6b,
title = "RANK receptor oligomerisation in the regulation of NFκB signalling",
abstract = "The interaction of receptor activator of NFκB (RANK), a member of the tumour necrosis factor receptor superfamily, with RANK ligand is crucial for the formation, function and survival of osteoclasts. The role of the cytoplasmic oligomerisation domain (pre-ligand assembly domain; PLAD or 'IVVY' motif) in the ligand-dependent activation of downstream NFκB signalling has not been studied previously. The discovery of truncating mutations of TNFRSF11A (W434X and G280X that lack the PLAD) as the cause of rare cases of osteoclast-poor osteopetrosis offered the opportunity for functional study of this region. Recapitulating the W434X mutation by transcription activator-like effector nuclease (TALEN)-mediated targeted disruption of Tnfrsf11a within the region homologous to W434X in the mouse macrophage-like cell line RAW264.7 impaired formation of osteoclast-like cells. Using overexpression studies, we demonstrated that, in contrast to WT-RANK, the absence of the PLAD in G280X-RANK and W434X-RANK prevented ligand-independent but not ligand-dependent oligomerisation. Cells expressing W434X-RANK, in which only two of the three TRAF6-binding motifs are present, continued to exhibit ligand-dependent NFκB signalling. Hence, the absence of the PLAD did not prevent ligand-induced trimerisation and subsequent NFκB activation of RANK, demonstrating that therapeutic targeting of the PLAD in the prevention of osteoporosis may not be as effective as proposed previously.",
keywords = "oligomerisation , RANK, NFκB, osteoperosis",
author = "S Das and I Sepahi and A Duthie and S Clark and Crockett, {J C}",
note = "{\circledC} 2014 The authors.",
year = "2014",
month = "8",
day = "1",
doi = "10.1530/JME-14-0077",
language = "English",
volume = "53",
pages = "81--91",
journal = "Journal of Molecular Endocrinology",
issn = "0952-5041",
publisher = "Society for Endocrinology",
number = "1",

}

TY - JOUR

T1 - RANK receptor oligomerisation in the regulation of NFκB signalling

AU - Das, S

AU - Sepahi, I

AU - Duthie, A

AU - Clark, S

AU - Crockett, J C

N1 - © 2014 The authors.

PY - 2014/8/1

Y1 - 2014/8/1

N2 - The interaction of receptor activator of NFκB (RANK), a member of the tumour necrosis factor receptor superfamily, with RANK ligand is crucial for the formation, function and survival of osteoclasts. The role of the cytoplasmic oligomerisation domain (pre-ligand assembly domain; PLAD or 'IVVY' motif) in the ligand-dependent activation of downstream NFκB signalling has not been studied previously. The discovery of truncating mutations of TNFRSF11A (W434X and G280X that lack the PLAD) as the cause of rare cases of osteoclast-poor osteopetrosis offered the opportunity for functional study of this region. Recapitulating the W434X mutation by transcription activator-like effector nuclease (TALEN)-mediated targeted disruption of Tnfrsf11a within the region homologous to W434X in the mouse macrophage-like cell line RAW264.7 impaired formation of osteoclast-like cells. Using overexpression studies, we demonstrated that, in contrast to WT-RANK, the absence of the PLAD in G280X-RANK and W434X-RANK prevented ligand-independent but not ligand-dependent oligomerisation. Cells expressing W434X-RANK, in which only two of the three TRAF6-binding motifs are present, continued to exhibit ligand-dependent NFκB signalling. Hence, the absence of the PLAD did not prevent ligand-induced trimerisation and subsequent NFκB activation of RANK, demonstrating that therapeutic targeting of the PLAD in the prevention of osteoporosis may not be as effective as proposed previously.

AB - The interaction of receptor activator of NFκB (RANK), a member of the tumour necrosis factor receptor superfamily, with RANK ligand is crucial for the formation, function and survival of osteoclasts. The role of the cytoplasmic oligomerisation domain (pre-ligand assembly domain; PLAD or 'IVVY' motif) in the ligand-dependent activation of downstream NFκB signalling has not been studied previously. The discovery of truncating mutations of TNFRSF11A (W434X and G280X that lack the PLAD) as the cause of rare cases of osteoclast-poor osteopetrosis offered the opportunity for functional study of this region. Recapitulating the W434X mutation by transcription activator-like effector nuclease (TALEN)-mediated targeted disruption of Tnfrsf11a within the region homologous to W434X in the mouse macrophage-like cell line RAW264.7 impaired formation of osteoclast-like cells. Using overexpression studies, we demonstrated that, in contrast to WT-RANK, the absence of the PLAD in G280X-RANK and W434X-RANK prevented ligand-independent but not ligand-dependent oligomerisation. Cells expressing W434X-RANK, in which only two of the three TRAF6-binding motifs are present, continued to exhibit ligand-dependent NFκB signalling. Hence, the absence of the PLAD did not prevent ligand-induced trimerisation and subsequent NFκB activation of RANK, demonstrating that therapeutic targeting of the PLAD in the prevention of osteoporosis may not be as effective as proposed previously.

KW - oligomerisation

KW - RANK

KW - NFκB

KW - osteoperosis

U2 - 10.1530/JME-14-0077

DO - 10.1530/JME-14-0077

M3 - Article

VL - 53

SP - 81

EP - 91

JO - Journal of Molecular Endocrinology

JF - Journal of Molecular Endocrinology

SN - 0952-5041

IS - 1

ER -