ICAP-1 monoubiquitylation coordinates matrix density and rigidity sensing for cell migration through ROCK2–MRCKα balance

Bouin, Anne-Pascale; Kyumurkov, Alexander; Régent-Kloeckner, Myriam; Ribba, Anne-Sophie; Faurobert, Eva; Fournier, Henri-Noël; Bourrin-Reynard, Ingrid; Manet-Dupé, Sandra; Oddou, Christiane; Balland, Martial; Planus, Emmanuelle; Albiges-Rizo, Corinne

doi:10.1242/jcs.200139

Cited by 8 publications

(10 citation statements)

References 39 publications

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“…Previously known as a regulator of cell migration, 33,34 and cell‐shape related cytoskeletal reorganization 35 (such as polarization, 36,37 protrusion, 38,39 and extrusion 40 ) in neurons, epithelial cells and osteoblasts, we show here the presence of MRCKα in the EPOR‐TFR2 complex in erythroblasts, and that MRCKα is required for efficient erythropoiesis. However, the mechanism by which MRCKα regulates erythropoiesis remains elusive.…”

Section: Discussionsupporting

confidence: 58%

Myotonic dystrophy kinase‐related CDC42‐binding kinase α, a new transferrin receptor type 2‐binding partner, is a regulator of erythropoiesis

et al. 2021

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Efficient erythropoiesis relies on the expression of the transferrin receptor type 2 (TFR2). In erythroid precursors, TFR2 facilitates the export of the erythropoietin receptor (EPOR) to cell surface, which ensures the survival and proliferation of erythroblasts. Although TFR2 has a crucial role in erythropoiesis regulation, its mechanism of action remains to be clarified. To understand its role better, we aimed at identifying its protein partners by mass‐spectrometry after immunoprecipitation in erythroid cells. Here we report the kinase MRCKα (myotonic dystrophy kinase‐related CDC42‐binding kinase α) as a new partner of both TFR2 and EPOR in erythroblasts. We show that MRCKα is co‐expressed with TFR2, and TFR1 during terminal differentiation and regulates the internalization of the two types of transferrin receptors. The knockdown of MRCKα by shRNA in human primary erythroblasts leads to a decreased cell surface expression of both TFR1 and TFR2, an increased cell‐surface expression of EPOR, and a delayed differentiation. Additionally, knockout of Mrckα in the murine MEDEP cells also leads to a striking delay in erythropoiesis, showcasing the importance of this kinase in both species. Our data highlight the importance of MRCKα in the regulation of erythropoiesis.

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Section: Discussionsupporting

confidence: 58%

Myotonic dystrophy kinase‐related CDC42‐binding kinase α, a new transferrin receptor type 2‐binding partner, is a regulator of erythropoiesis

et al. 2021

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“…Our present results implicate a critical contribution of the integrin inhibitor, ICAP-1, in cellular mechanotransduction by working in concert with NME2 NDPK to control clathrin-mediated endocytosis of integrins at the edge of FAs. This new mechanism illustrates the importance of integrin inhibitors in mechanosensing and mechanoresponsiveness (Bouin et al, 2017;Lerche et al, 2020). Mechanistically, ICAP-1 acts as an adaptor protein in integrin endocytosis.…”

Section: Discussionmentioning

confidence: 74%

“…S1, D and E ), we checked that the combined deletion of β1 integrin and ICAP-1 did not affect the total expression of β3 integrin. Cells were seeded on Fn-coated polyacrylamide hydrogels (PA hydro gels) with a Young’s modulus (E) of 5 kPa as previously described ( Bouin et al, 2017 ). First, we observed that osteoblasts lacking β1 integrin ( β1 integrin –/– -icap-1 +/+ ) were defective in force generation ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We have shown that ICAP-1 is involved in cell mechanical properties and cell differentiation in a β1 integrin-dependent manner ( Brunner et al, 2011 ; Faurobert et al, 2013 ; Bouvard et al, 2007 ; Renz et al, 2015 ; Millon-Frémillon et al, 2008 ). We have also proposed ICAP-1 as a key modulator of cellular mechanoresponse in a β1-integrin–independent manner ( Bouin et al, 2017 ). To delve deeper into the ICAP-1 functional properties, we interrogated β1-integrin–dependent and –independent contributions of ICAP-1 to the cell mechanical response.…”

Section: Introductionmentioning

confidence: 99%

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Force tuning through regulation of clathrin-dependent integrin endocytosis

Kyumurkov

Bouin

Boissan

et al. 2022

Journal of Cell Biology

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Integrin endocytosis is essential for many fundamental cellular processes. Whether and how the internalization impacts cellular mechanics remains elusive. Whereas previous studies reported the contribution of the integrin activator, talin, in force development, the involvement of inhibitors is less documented. We identified ICAP-1 as an integrin inhibitor involved in mechanotransduction by co-working with NME2 to control clathrin-mediated endocytosis of integrins at the edge of focal adhesions (FA). Loss of ICAP-1 enables β3-integrin-mediated force generation independently of β1 integrin. β3-integrin-mediated forces were associated with a decrease in β3 integrin dynamics stemming from their reduced diffusion within adhesion sites and slow turnover of FA. The decrease in β3 integrin dynamics correlated with a defect in integrin endocytosis. ICAP-1 acts as an adaptor for clathrin-dependent endocytosis of integrins. ICAP-1 controls integrin endocytosis by interacting with NME2, a key regulator of dynamin-dependent clathrin-coated pits fission. Control of clathrin-mediated integrin endocytosis by an inhibitor is an unprecedented mechanism to tune forces at FA.

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“…Consequently, their in vitro data might point towards a certain resistance to ROCK inhibition in EWS cells. Of note, it is well established that >1 µM SR3677 acts on several off-target kinases ( 33 ) including PKA [which promotes tumor growth and metastasis in EWS ( 37 ), MRCK [another mediator of cell contractibility ( 38 ) and AKT1 that plays important roles in EWS survival ( 39 ).…”

Section: Discussionmentioning

confidence: 99%

Prognostic value and functional role of ROCK2 in pediatric Ewing sarcoma

et al. 2017

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Ewing's sarcoma (EWS) is a highly aggressive bone cancer that affects children and adolescents. Despite advances in multimodal management, 5-year event-free survival rates for patients presenting with metastases at diagnosis remain at 25%. As key regulators of actin organization, the Rho-associated coiled-coil containing protein kinases, ROCK1 and ROCK2, have been associated with cancer dissemination and poorer prognosis. Recently, in vitro data indicating ROCK2 as a molecular target for the treatment of EWS has been presented. Nonetheless, a deeper exploration of the contribution of this kinase dysregulation in EWS is still necessary. In this regard, the present study aimed to evaluate the expression of ROCK1 and ROCK2 in 23 pediatric tumor samples and to verify the prospect of using their pharmacological inhibition through functional assays. Our results showed positive immunostaining for ROCK1 and ROCK2 in the majority samples (75 and 65%, respectively). A significantly increased risk of incomplete remission in patients with positive immunostaining for ROCK2 was found (P=0.026), though no correlations with other prognostic features (huvos classification, FLI1/EWS status, relapse, metastasis or death) were observed. Associations with survival were merely suggestive. Apparent protein expression of both kinases was also found in EWS cell lines (SK-ES-1 and RD-ES). Treatments with selective ROCK inhibitors did not alter cell viability or migration in vitro. However, a significant increase in invasion was observed after treatment with SR3677 (ROCK2 inhibitor) and hydroxyfasudil (pan-inhibitor). Consequently, even though the majority of EWS samples included in our study showed positivity for ROCK1 and ROCK2, the lack of significant associations with prognosis and absence of appropriate responses to their inhibition in vitro does not support their prospective use as therapeutic targets for the treatment of this metastatic tumor. Larger cohort studies might provide more evidence on whether there is a specific role of ROCK kinases in EWS physiopathology.

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ICAP-1 monoubiquitylation coordinates matrix density and rigidity sensing for cell migration through ROCK2–MRCKα balance

Cited by 8 publications

References 39 publications

Myotonic dystrophy kinase‐related CDC42‐binding kinase α, a new transferrin receptor type 2‐binding partner, is a regulator of erythropoiesis

Myotonic dystrophy kinase‐related CDC42‐binding kinase α, a new transferrin receptor type 2‐binding partner, is a regulator of erythropoiesis

Force tuning through regulation of clathrin-dependent integrin endocytosis

Prognostic value and functional role of ROCK2 in pediatric Ewing sarcoma

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