Cytoplasmic sequestration of the RhoA effector mDiaphanous1 by Prohibitin2 promotes muscle differentiation

Abstract

Running title: Phb2 interacts with mDia1 in myotubesSummary statement: mDia1 has common and stage-specific functions in muscle cells. In myotubes, mDia1 is sequestered by an interacting protein Prohibitin2, which promotes Myogenin expression and mitigates mDia1's inhibitory effects on differentiation. (which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint . http://dx.doi.org/10.1101/283044 doi: bioRxiv preprint first posted online Mar. 15, 2018; 2 Abstract Adhesion and growth factor dependent signalling control muscle gene expression through common effectors, coupling cytoskeletal dynamics to transcriptional activation.Earlier, we showed that mDiaphanous1, an effector of adhesion-dependent RhoA-signalling promotes MyoD expression in myoblasts, linking contractility to lineage determination. Here, we report that paradoxically, mDia1 negatively regulates MyoD function in myotubes.Knockdown of endogenous mDia1 during differentiation enhances MyoD and Myogenin expression, while over-expression of mDia1ΔN3, a RhoA-independent mutant, suppresses Myogenin promoter activity and expression. We investigated mechanisms that may counteract mDia1 to promote Myogenin expression and timely differentiation by analysing mDia1-interacting proteins. We report that mDia1 has a stage-specific interactome, including Prohibitin2, MyoD, Akt2, and β-Catenin, of which Prohibitin2 colocalises with mDia1 in cytoplasmic punctae and opposes mDia1 function in myotubes. Co-expression of mDia1-binding domains of Prohibitin2 reverses the anti-myogenic effects of mDia1ΔN3. Our results suggest that Prohibitin2 sequesters mDiaphanous1 to dampen its activity and finetune RhoAmDiaphanous1 signalling to promote differentiation. Overall, we report that mDia1 is multifunctional signaling effector with opposing functions in different cellular stages, but is modulated by a differentiation-dependent interactome.All rights reserved. No reuse allowed without permission.(which was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.

Mitochondria–actin cytoskeleton crosstalk in cell migration

“…Since formins also interact with microtubules, long‐distance mitochondrial transport is also sensitive to perturbation of formins (Gaillard et al, 2011). Mass‐spectrometric analysis has shown that mDia1 interacts with several components of the mitochondrial OxPhos complexes‐I, ‐II, ‐III, ‐IV, and ‐V, and influences mitochondrial biogenesis (Saleh et al, 2019). Although mDia1 strongly associates with prohibitin 2 (Phb2—an important regulator of mitochondrial bioenergetics) (Signorile et al, 2019), the experimental evidence for mDia1's involvement in mitochondrial OxPhos regulation is currently lacking in the literature.…”