Integral Membrane Protein 2A Is a Negative Regulator of Canonical and Non-Canonical Hedgehog Signalling.

Morales-Alcala, Cintli C, Georgiou, Ioanna Ch ORCID: 0000-0003-3055-1105, Timmis, Alex J and Riobo-Del Galdo, Natalia A ORCID: 0000-0002-8942-7873 (2021) Integral Membrane Protein 2A Is a Negative Regulator of Canonical and Non-Canonical Hedgehog Signalling. Cells, 10 (8). 2003-.

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Abstract

The Hedgehog (Hh) receptor PTCH1 and the integral membrane protein 2A (ITM2A) inhibit autophagy by reducing autolysosome formation. In this study, we demonstrate that ITM2A physically interacts with PTCH1; however, the two proteins inhibit autophagic flux independently, since silencing of ITM2A did not prevent the accumulation of LC3BII and p62 in PTCH1-overexpressing cells, suggesting that they provide alternative modes to limit autophagy. Knockdown of ITM2A potentiated PTCH1-induced autophagic flux blockade and increased PTCH1 expression, while ITM2A overexpression reduced PTCH1 protein levels, indicating that it is a negative regulator of PTCH1 non-canonical signalling. Our study also revealed that endogenous ITM2A is necessary for timely induction of myogenic differentiation markers in C2C12 cells since partial knockdown delays the timing of differentiation. We also found that basal autophagic flux decreases during myogenic differentiation at the same time that ITM2A expression increases. Given that canonical Hh signalling prevents myogenic differentiation, we investigated the effect of ITM2A on canonical Hh signalling using GLI-luciferase assays. Our findings demonstrate that ITM2A is a strong negative regulator of GLI transcriptional activity and of GLI1 stability. In summary, ITM2A negatively regulates canonical and non-canonical Hh signalling.

Item Type:	Article
Uncontrolled Keywords:	Hela Cells, NIH 3T3 Cells, Myoblasts, Skeletal, Animals, Humans, Mice, Membrane Proteins, Signal Transduction, Cell Differentiation, Protein Binding, Muscle Development, Autophagy, HEK293 Cells, Protein Interaction Maps, Zinc Finger Protein GLI1, Patched-1 Receptor, Autophagy-Related Proteins
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User:	Symplectic Admin
Date Deposited:	04 Mar 2024 09:10
Last Modified:	04 Mar 2024 09:10
DOI:	10.3390/cells10082003
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3179060