A Dock-and-Lock Mechanism Clusters ADAM10 at Cell-Cell Junctions to Promote α-Toxin Cytotoxicity

Shah, Jimit; Rouaud, Florian; Guerrera, Diego; Vasileva, Ekaterina; Popov, Lauren M.; Kelley, William L.; Rubinstein, Eric; Carette, Jan E.; Amieva, Manuel R.; Citi, Sandra

doi:10.1016/j.celrep.2018.10.088

Cited by 43 publications

(77 citation statements)

References 64 publications

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“…Thus, Tspan15 and ADAM10 appear to be each required for the normal expression of the other. Indeed, it is well established that TspanC8s are important for ADAM10 maturation and trafficking to the cell surface (Dornier et al 2012; Haining et al 2012; Prox et al 2012; Zhou et al 2014; Jouannet et al 2016; Noy et al 2016; Virreira Winter et al 2016; Reyat et al 2017; Brummer et al 2018; Seipold et al 2018; Shah et al 2018; Brummer et al 2019), and we extended these observations by showing that ADAM10 surface expression was partially reduced in the absence of Tspan15 in A549, HEK-293T and HUVECs. Expression of other TspanC8s by these cells (Haining et al 2012; Matthews et al 2017) is likely to be the reason that the reduction is only partial.…”

Section: Discussionsupporting

confidence: 66%

Tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Koo

Harrison

Noy

et al. 2019

Preprint

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1A disintegrin and metalloprotease 10 (ADAM10) is essential for embryonic development and 2 impacts on diseases such as cancer, Alzheimer's and inflammatory diseases. ADAM10 is a 3 'molecular scissor' that proteolytically cleaves the extracellular region from over 100 substrates, 4 including Notch, amyloid precursor protein, cadherins, growth factors and chemokines. 5 ADAM10 was recently proposed to function as six distinct scissors with different substrates, 6 depending on its association with one of six regulatory tetraspanins, termed TspanC8s. 7 However, it remains unclear to what degree ADAM10 function is critically dependent on a 8 TspanC8 partner. To address this, we generated the first monoclonal antibodies to Tspan15 as a 9 model TspanC8. These were used to show that ADAM10 is the principal Tspan15-interacting 10 protein, that Tspan15 expression requires ADAM10 in cell lines and primary cells, and that a 11 synthetic ADAM10/Tspan15 fusion protein is a functional scissor. Together these findings 12 suggest that ADAM10 exists as an intimate ADAM10/TspanC8 scissor complex. 13 14 15A disintegrin and metalloproteinase 10 (ADAM10) is a ubiquitously-expressed transmembrane 16 protein which acts as a 'molecular scissor' by cleaving the extracellular region from over 100 17 substrates, a process termed ectodomain shedding (Lichtenthaler et al. 2018). ADAM10 is 18 essential for embryonic development by activating Notch proteins which determine cell fate. 19 Other substrates include cadherin adhesion molecules, amyloid precursor protein, and 20 transmembrane growth factors and chemokines. As such, ADAM10 is important for health and 21 in diseases such as cancer, Alzheimer's and inflammatory diseases (Wetzel et al. 2017). 22 The tetraspanins are a superfamily of 33 transmembrane proteins in mammals which interact 23 with specific transmembrane partner proteins and regulate their intracellular trafficking, lateral 24 mobility and clustering at the cell surface (Termini and Gillette 2017; van Deventer et al. 2017). 25 42 Jouannet et al. 2016; Noy et al. 2016; Seipold et al. 2018). Tspan15 is also upregulated, and is a 43 marker of poor prognosis, in certain cancers (Zhang et al. 2018; Hiroshima et al. 2019; 44 Sidahmed-Adrar et al. 2019) and promotes cancer progression in a mouse model (Zhang et al. 45 2018). The aims of this study were to generate the first Tspan15 mAbs and to test three 46 hypotheses that would support the theory that Tspan15 and ADAM10 exist together as a scissor 47 complex: first, that ADAM10 is the principal Tspan15-interacting protein; second, that Tspan15 48 expression requires ADAM10; and third, that covalently linking Tspan15 and ADAM10 together 49 as a single fusion protein yields a functional scissor. 50 51 Results 52Generation of Tspan15 mAbs 53 The majority of anti-tetraspanin mAbs have epitopes within the large extracellular loop (LEL). 54 However, it has been traditionally difficult to make mAbs to many tetraspanins due to lack of 55 efficacy of recombinant LELs a...

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Section: Discussionsupporting

confidence: 66%

Tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Koo

Harrison

Noy

et al. 2019

Preprint

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“…By GST pulldown analysis, the N-terminal region of PLEKHA7 binds weakly to the C-terminal region of Tspan33, but this interaction is greatly enhanced in the presence of PDZD11 (13). In agreement, PDZD11 is required in cells for the interaction of PLEKHA7 with Tspan33, and clustering of the Tspan33-ADAM10 complex at junctions (13).…”

Section: Pdzd11 Promotes the Interaction Of Tspan33 With Plekha7 By Imentioning

confidence: 75%

Cooperative binding of the tandem WW domains of PLEKHA7 to PDZD11 promotes conformation-dependent interaction with tetraspanin 33

Rouaud

Tessaro

Aimaretti

et al. 2020

Journal of Biological Chemistry

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Pleckstrin homology domain–containing A7 (PLEKHA7) is a cytoplasmic protein at adherens junctions that has been implicated in hypertension, glaucoma, and responses to Staphylococcus aureus α-toxin. Complex formation between PLEKHA7, PDZ domain–containing 11 (PDZD11), tetraspanin 33, and the α-toxin receptor ADAM metallopeptidase domain 10 (ADAM10) promotes junctional clustering of ADAM10 and α-toxin–mediated pore formation. However, how the N-terminal region of PDZD11 interacts with the N-terminal tandem WW domains of PLEKHA7 and how this interaction promotes tetraspanin 33 binding to the WW1 domain is unclear. Here, we used site-directed mutagenesis, glutathione S-transferase pulldown experiments, immunofluorescence, molecular modeling, and docking experiments to characterize the mechanisms driving these interactions. We found that Asp-30 of WW1 and His-75 of WW2 interact through a hydrogen bond and, together with Thr-35 of WW1, form a binding pocket that accommodates a polyproline stretch within the N-terminal PDZD11 region. By strengthening the interactions of the ternary complex, the WW2 domain stabilized the WW1 domain and cooperatively promoted the interaction with PDZD11. Modeling results indicated that, in turn, PDZD11 binding induces a conformational rearrangement, which strengthens the ternary complex, and contributes to enlarging a “hydrophobic hot spot” region on the WW1 domain. The last two lipophilic residues of tetraspanin 33, Trp-283 and Tyr-282, were required for its interaction with PLEKHA7. Docking of the tetraspanin 33 C terminus revealed that it fits into the hydrophobic hot spot region of the accessible surface of WW1. We conclude that communication between the two tandem WW domains of PLEKHA7 and the PLEKHA7–PDZD11 interaction modulate the ligand-binding properties of PLEKHA7.

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“…ADAM10 is a transmembrane protease that cleaves extracellular regions of several transmembrane target proteins, including Notch . Furthermore, other Tspans have been shown to regulate ADAM10 maturation and activity, and Tspan3 itself has been shown to regulate ADAM10 expression . Our studies show that the Cldn‐11 effect on Notch signaling may be mediated via Tspan3 regulation of ADAM10 function.…”

Section: Discussionmentioning

confidence: 66%

“…ADAM10 is a transmembrane protein that cleaves extracellular regions of several transmembrane target proteins, including Notch, and we previously showed that osteoblasts express ADAM10 . Further, it was recently found that mutation of Tspan14 reduced ADAM10 surface expression and activity in human umbilical vein endothelial cells, and a number of other Tspans, including Tspan5, Tspan10, Tspan15, Tspan17, and Tspan33, have been shown to regulate ADAM10 activity . Notably, Tspan3 has also been found to regulate ADAM10 expression .…”

Section: Resultsmentioning

confidence: 93%

Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling

Lindsey

Xing

Pourteymoor

et al. 2019

Journal of Bone and Mineral Research

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The claudin (Cldn) family comprises 27 members of 20 to 34 kDa transmembrane tight junction proteins. In addition to Cldns' established canonical role as barriers controlling paracellular flow of molecules, a distinct noncanonical role for them as mediators of cell signaling is now emerging. In our studies evaluating Cldn family expression levels during osteoblast differentiation, Cldn-11 showed the largest increase (60-fold). Immunohistochemistry studies revealed high Cldn-11 expression in trabecular (Tb) bone lining cells. Micro-CT analysis of femurs and vertebrae of Cldn-11 knock-out (KO) mice at 12 weeks of age exhibited a 40% (p < 0.01) reduction in Tb bone volume adjusted for tissue volume compared with control mice, a change caused by significant reductions in Tb number and thickness and increase in Tb separation. Histomorphometry and serum biomarker studies revealed that reduced bone formation, not increased resorption, is the cause for reduced Tb bone volume in the Cldn-11 KO mice. Cldn-11 KO osteoblasts expressed reduced ALP and BSP, whereas Cldn-11 overexpression in MC3T3-E1 cells increased expression of ALP and BSP. Mechanistically, Cldn-11 interacted with tetraspanin (Tspan)3 in osteoblasts, and Tspan3 knockdown reduced osteoblast differentiation. Because members of the Tspan family regulate cell functions via Notch signaling, we evaluated whether Cldn-11/ Tspan3 regulates Notch signaling in osteoblasts. Accordingly, Notch targets Hey1 and Hey2 were significantly upregulated in Cldn-11 overexpressing cultures but downregulated in both Cldn-11 KO and Tspan3 knockdown osteoblasts. Because ADAM10 has been shown to interact with Tspan family members to regulate Notch signaling, we evaluated whether Cldn-11 regulates ADAM10 expression. Cldn-11 overexpressing cells express more mature ADAM10, and an ADAM10 inhibitor blocked the Cldn-11 effect on osteoblast differentiation. Based on these data, we propose Cldn-11 as a novel component of an osteoblast cell surface protein complex, comprising Tspan3 and ADAM10, which regulates Notch signaling and cell differentiation.

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A Dock-and-Lock Mechanism Clusters ADAM10 at Cell-Cell Junctions to Promote α-Toxin Cytotoxicity

Cited by 43 publications

References 64 publications

Tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex

Cooperative binding of the tandem WW domains of PLEKHA7 to PDZD11 promotes conformation-dependent interaction with tetraspanin 33

Novel Role for Claudin-11 in the Regulation of Osteoblasts via Modulation of ADAM10-Mediated Notch Signaling

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