Professor Steve Winder
Professor of Molecular Cell Biology
Director of Postgraduate Teaching
Department of Biomedical Science
The University of Sheffield
Sheffield S10 2TN
Tel: +44 (0) 114 222 2332
Room: B2 06 Florey building
The main research focus of my group is centered around understanding the normal functions of the cell adhesion and signalling adaptor protein dystroglycan and in diseases such as muscular dystrophy and cancer. In particular we are investigating the regulation of the actin cytoskeleton, adhesion mediated signalling and cell motility and invasion using molecular cell biology and transgenic approaches.
My research group is part of the Centre for Membrane Interactions and Dynamics (CMIAD).
Activities and distinctions
Selected publications since 2014
The laminin binding protein dystroglycan plays multiple roles in cell adhesion, signalling and membrane cytoskeleton stability. Perturbation of dystroglycan function underlies several muscular dystrophies and is also a secondary consequence of adenocarcinoma progression. Changes to the post-translational modification of dystroglycan are crucial in directing the associations, cellular localisation and ultimately degradation of dystroglycan. Our aim is to elucidate the mechanisms and consequences of these post-translational modifications in order to better understand dystroglycan function and to identify potential therapeutic targets for the treatment of muscular dystrophy or cancer.
We employ in vitro, in/ex vivo fish and mouse genetic models with clinically relevant archival tissue samples or immortalised cell lines. Dystroglycan function is dissected through the use of molecular cell biology approaches, and potential therapeutic targets are assessed in vitro and in vivo. Recently we have developed a novel therapeutic approach for the treatment of Duchenne muscular dystrophy using inhibitors of tyrosine phosphorylation and proteasomal degradation. Through the use of zebrafish screening and phenotypic analysis in mdx mice and human DMD myoblasts we are in the process of validating the potential for repurposed drugs as a precursor to initiating clinical trials. Physiological analysis is carried out I collaboration with Nic Wells at the RVC London.
In vitro models of prostate cancer have revealed a role for the post-translational proteolytic processing and nuclear targeting of dystroglycan. Current efforts are centred around characterising a role as part of the LINC complex in the inner nuclear membrane. These studies form part of an ongoing collaboration with Bulmaro Cisneros, CINVESTAV Mexico City.
Postgraduate PhD Opportunities
1. The Dystroglycan LINC: the functions of dystroglycan in the nuclear envelope in muscular dystrophy and cancer
Co-supervisor: Professor Jamie Hobbs (Physics and Astronomy)
Biological membranes in animals have little intrinsic resistance to mechanical forces. Instead support is provided by membrane associated protein complexes. The plasma membrane, mitochondrial membrane and nuclear membrane all possess distinct integral membrane protein complexes that connect to the cytoskeleton and protect and position the membrane within the cell. We have found recently that dystroglycan, a plasma membrane adhesion receptor, characterised primarily for its role in membrane of striated muscle is also present in the in the nuclear envelope. Moreover, we have found that dystroglycan depletion leads to disruption of the nuclear membrane.
Novelty and Timeliness
Dystroglycan is the only example of a transmembrane adhesion receptor that has structural and mechanical roles in plasma membrane and nuclear membrane. Through cytoskeletal connections it provides support to both membrane structures in tissues like muscle that are subjected to continued physical stress. The Winder lab and collaborators in Mexico are the only groups worldwide working on this novel aspect of dystroglycan function.
This is an integrated multidisciplinary project involving molecular cell biology, biophysics and biomechanics. The student will use these approaches in normal and dystroglycan knock-out myoblasts, including morphological analysis by immunofluorescence and analysis of interactions by immunoprecipitation. Migration of cells in 2D and 3D microfluidic chambers will assess the mechanical properties of the nucleus to deformation, whilst atomic force microscopy will assess the contribution of dystroglycan to the biophysical properties of isolated nuclei and nuclei in situ.
Dystroglycan is an essential protein that is a linker through biological membranes including the plasma membrane and the nuclear membrane. This project will address fundamental aspects of nuclear structure and dynamics employing a multidisciplinary approach comprising molecular cell biology and advanced microscopy. The project will be supervised by two world leading experts in their fields: Professor Steve Winder in the Department of Biomedical Science who works on the cell biology of dystroglycan, and Professor Jamie Hobbs who is at the forefront of research advances using atomic force microscopy (AFM) to probe biological systems.
The project will use genetic approaches in cells in tissue culture to manipulate the function of dystroglycan, a newly identified component of the nuclear membrane, and how it contributes to nuclear shape, nuclear resistance to stress and nuclear position in the cell. We will use biophysical approaches to measure nuclear deformation and stability and cell biological approaches to investigate the effects of altered nuclear deformation on cell migration in two and three dimensions, using microfluidic chambers. AFM and high resolution fluorescence microscopy, coupled with genetic manipulation will provide a powerful combination of approaches to understand the essential functions of dystroglycan in the nucleus.
2. Targeting dystroglycan to the nucleus in muscular dystrophy and cancer
Dystroglycan is an essential cell adhesion receptor required for early embryonic development. Genetic loss of function gives rise to severe muscular dystrophies with neuronal involvement. Post-translational loss of function also occurs in Duchenne muscular dystrophy and in some cancers. This includes phosphorylation, proteolysis and ubiquitination. Moreover some proteolytic fragments of dystroglycan are targeted to the nucleus where they have effects on transcription. As part of our analysis of dystroglycan post-translational modifications, we identified a lipid modification - palmitoylation of a conserved cysteine residue that could act to anchor bioactive dystroglycan fragments to the membrane, both at the cell surface and in the nucleus.
To examine the function of dystroglycan palmitoylation in cellular targeting and in cellular phenotypes associated with muscular dystrophy and cancer.
PCR-based site-directed mutagenesis, and cloning of dystroglycan mutants. Overexpression of dystroglycan mutants in tissue culture cells. Analysis of subcellular distribution of dystroglycan by quantitative immunofluorescence microscopy and cell fractionation. In vitro assays of cell invasion and metastatic growth.
For further information about these projects and how to apply, please see our PhD Opportunities page.
Miss Katherine Collins-Taylor
Ms Tracy Emmerson
Miss Laura Jacobs
Dr Gemma Woodward