3BP2 Adapter Protein Is Required for Receptor Activator of NFκB Ligand (RANKL)-induced Osteoclast Differentiation of RAW264.7 Cells

Guezguez, Amel; Prod’homme, Virginie; Mouska, Xavier; Baudot, Alice D.; Blin-Wakkach, Claudine; Rottapel, Robert; Deckert, Marcel

doi:10.1074/jbc.m109.091124

Cited by 35 publications

(39 citation statements)

References 52 publications

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“…It is important to note that defects observed in 3BP2-deficient osteoclast appearance and function in vitro were much more severe than those observed in vivo, suggesting that a factor or combination of factors present in vivo partially rescues osteoclast function in the absence of 3BP2. In contrast to our results obtained in primary bone marrow-derived cells, 3BP2 has recently been shown to be required in the monocytic cell line RAW264.7 for osteoclast differentiation and NFATc1 induction (48). The difference in our findings may be a reflection of the differential signaling requirements characteristic for this cell line compared with primary monocytes.…”

Section: Discussioncontrasting

confidence: 56%

3BP2-deficient mice are osteoporotic with impaired osteoblast and osteoclast functions

Levaot¹,

Simoncic²,

Dimitriou³

et al. 2011

J. Clin. Invest.

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A fine balance between bone resorption by osteoclasts and bone formation by osteoblasts maintains bone homeostasis. In patients with cherubism, gain-of-function mutations in 3BP2, which is encoded by SH3-domain binding protein 2 (SH3BP2), cause cystic lesions with activated osteoclasts that lead to craniofacial abnormalities. However, little is known about the function of wild-type 3BP2 in regulating bone homeostasis. Here we have shown that 3BP2 is required for the normal function of both osteoblasts and osteoclasts. Initial analysis showed that Sh3bp2 -/-mice developed osteoporosis as a result of reduced bone formation despite the fact that bone resorption was impaired. We demonstrated using reciprocal bone marrow chimeras, a cellintrinsic defect of the osteoblast and osteoclast compartments in vivo. Further, Sh3bp2 -/-osteoblasts failed to mature and form mineralized nodules in vitro, while Sh3bp2 -/-osteoclasts spread poorly and were unable to effectively degrade dentine matrix in vitro. Finally, we showed that 3BP2 was required for Abl activation in osteoblasts and Src activation in osteoclasts, and demonstrated that the in vitro defect of each cell type was restored by the respective expression of activated forms of these kinases. These findings reveal an unanticipated role for the 3BP2 adapter protein in osteoblast function and in coordinating bone homeostatic signals in both osteoclast and osteoblast lineages. Introduction3BP2 is an adapter protein that contains an N-terminal pleckstrin homology (PH) domain, a proline-rich stretch that binds to Src homology 3 (SH3) domain-containing proteins, and a C-terminal SH2 domain that binds to phosphotyrosine residues (1). 3BP2 was initially identified as a binding protein of the tyrosine kinase Abl (2). Work from our lab and others has identified the Src family kinases (SFKs), Syk, and the Vav family of Rho guanine nucleotide exchange factors (GEFs) as 3BP2-binding partners (1), all of which are known to play important roles in osteoclast function (3-5). Cherubism is a dominantly inherited syndrome characterized by excessive maxillary and mandibular bone resorption that is associated with activated osteoclasts and inflammatory cells creating interosseous cystic lesions (6). Single missense mutations in the gene encoding the adapter protein 3BP2 result in a gain-of-function alteration in the protein and is associated with the majority of cherubism patients (7). A mouse model that harbors 2 copies of a cherubism allele develops severe osteoporosis associated with highly activated osteoclasts (8).In order to elucidate the role of the wild-type form of 3BP2 in bone homeostasis, we analyzed loss-of-function mutant mice. As distinct from the osteoporotic phenotype of mice expressing the cherubism gain-of-function form of 3BP2 associated with active osteoclasts, we uncovered a complex bone phenotype in mice lacking 3BP2 characterized by loss-of-function in both the osteoclast and osteoblast lineages resulting in net decreased bone mineral density and reduced mechanical ...

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Section: Discussioncontrasting

confidence: 56%

3BP2-deficient mice are osteoporotic with impaired osteoblast and osteoclast functions

Levaot¹,

Simoncic²,

Dimitriou³

et al. 2011

J. Clin. Invest.

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“…Conversely, phagocytic capacity of heterozygous cherubic BMM was exacerbated compared with that of WT macrophages ( Figure 2D). TLR and Fc receptors signal via different cytosolic molecular pathways (14,15), and 3BP2 has been implicated in a variety of membrane receptor signaling pathways, including those stimulated by immunoreceptors (4, 5), RANK (8), and GPCR (16). Therefore, it seems unlikely that 3BP2 interacts with specific regulators of either TLR or Fc receptor pathways in macrophages.…”

Section: Wt/kimentioning

confidence: 99%

“…*P < 0.05; **P < 0.01; ***P < 0.001. Alternatively, polymerized actin was analyzed using a Zeiss Axiovert fluorescence microscope, as described before (8). RAC activities were measured on BMM lysates using G-LISA RAC Activation Assays (Cytoskeleton).…”

Section: Sh3bp2mentioning

confidence: 99%

“…Immunoblotting and co-IP were performed on BMM lysates as described (8). Abs were as follows: rabbit and goat Abs against 3BP2 (5,8); Abs against phosphorylated forms of SRC, SYK, ERK1/2, p38, JNK1/2, AKT, IKKα/β, IκBα (Cell Signaling Technology); anti-p-VAV1, and ERK2 (Santa Cruz Biotechnology Inc.).…”

Section: Sh3bp2mentioning

confidence: 99%

“…The most frequent SH3BP2 mutations affect exon 9, leading to the substitution of amino acids Arg415, Pro418, or Gly420 within an RSPPDG motif of the protein 3BP2 (1)(2)(3). 3BP2 is a cytoplasmic adapter involved in leukocyte activation (4)(5)(6)(7), osteoclast differentiation, and bone remodeling (8,9). 3BP2 signals downstream of diverse leukocyte membrane receptors through interactions with SRC and SYK kinases, PLCγ, and VAV GEF (2,3).…”

Section: Introductionmentioning

confidence: 99%

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Cherubism allele heterozygosity amplifies microbe-induced inflammatory responses in murine macrophages

Prod’homme¹,

Boyer²,

Dubois³

et al. 2015

J. Clin. Invest.

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Molecular Genetics of Cherubism

Reichenberger

Ueki

2016

Encyclopedia of Life Sciences

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Cherubism is a rare fibro‐osseous genetic disorder caused and maintained by autoinflammatory processes that are caused by mutations in the SH3BP2 gene. Cherubism is a form of fibrodysplasia of the jaws that affects children at 2–6 years of age. The typical symmetrical bone resorption lacunae are self‐limiting to the jaws. Bone is replaced with fibrous tissue that can expand significantly and lead to the characteristic facial swelling. Disease progression normally stops at puberty and regresses thereafter. Mutations in SH3BP2 affect a six‐amino acid recognition sequence for tankyrase, a poly(ADP‐ribose)polymerase that marks proteins for ubiquitination and degradation. SH3BP2 has multiple functions in cell types of hematopoietic origin. Elevated SH3BP2 protein levels in macrophage/osteoclast precursors cause increased osteoclastogenesis with reduced thresholds for stimulants. Increased TNF‐α levels generate an inflammatory environment in a cherubism mouse model and conceivably also in the oral cavity of cherubism patients. More research to explain the spatiotemporal progression of cherubism is needed. Key Concepts Cherubism is a temporospatially restricted bone lysis disorder with bone being substituted by expansile fibrous tissue. Patients with cherubism develop symmetrical bone lesions only in the maxilla and mandible. Signs of cherubism develop in children as early as 2 years of age and bone lysis typically resolves after puberty. First diagnosed often on dental radiographs. Known mutations are in the adaptor protein SH3BP2, which is involved in regulating the adaptive and innate immune system. Mutations in SH3BP2 are within a six‐amino acid interval and cause inhibition of tankyrase binding, which is needed for efficient degradation. High levels of intracellular SH3BP2 protein cause increased osteoclastogenesis and bone resorption in jaws. Bone lysis is regulated via TLR2/TLR4 and TNF‐α‐dependent autoinflammatory response. Jaw‐specific bone resorption in cherubism patients likely involves response to oral bacteria (pathogen‐associated molecular patterns; PAMPS) and bone remodelling during tooth eruption (damage‐associated molecular patterns; DAMPS). Diagnosis for some patients is consistent with cherubism but no mutations are found in exons of SH3BP2 .

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3BP2 Adapter Protein Is Required for Receptor Activator of NFκB Ligand (RANKL)-induced Osteoclast Differentiation of RAW264.7 Cells

Cited by 35 publications

References 52 publications

3BP2-deficient mice are osteoporotic with impaired osteoblast and osteoclast functions

3BP2-deficient mice are osteoporotic with impaired osteoblast and osteoclast functions

Cherubism allele heterozygosity amplifies microbe-induced inflammatory responses in murine macrophages

Molecular Genetics of Cherubism

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