Brief career history
- 2016-present: Head of Department, Biomedical Science
- 2012-2016: Professor, Deputy Head of Biomedical Science
- 2008-2012: Professor and MRC Senior Research Fellow. Department of Molecular Biology and Biotechnology, University of Sheffield
- 2003–2008: MRC Senior Research Fellow. Department of Molecular Biology and Biotechnology, University of Sheffield
- 1999–2003: MRC Senior Research Fellow, Department of Biochemistry, University of Glasgow
- 1996–1999: Wellcome Trust Career Development Fellow, Department of Biochemistry, University of Dundee
- 1993-1999: Wellcome International Travelling Fellow, University of California, Berkeley, USA. Supervisor David Drubin
- 1989–1993: PhD Imperial Cancer Research Fund (now CRUK) and University College London. Supervisor Graham Warren
- 1985-1989: BA MA(Oxon) Biochemistry, University of Oxford. 1st Class.
The main research focus of my lab is to understand the mechanisms governing cell organization and architecture. We are currently addressing how one of the key elements of the cytoskeleton (actin) can initiate filaments formation and how these filaments can then be organized to allow inward bending of the plasma membrane. This invagination process is called endocytosis and allows cells to regulate their cell surface composition in response to environmental signals. We are also aiming to understand how the process of endocytosis is coupled to the invasion and survival properties of the human fungal pathogen Candida albicans.
- MRC Senior non-clinical Fellow (2001 – 2012)
- Editorial Board for the international journals Molecular Biology of the Cell, Cytoskeleton and F1000 Research
- Faculty of 1000 member (Cytoskeleton panel from 2010)
- Grant awarding panels: The Wellcome Trust (Molecules, Genes and Cells Committee) 2006-2009; Wolfson-Royal Society Infrastructure Awards (2012)
- Scientific Advisory Board, Biological Sciences, Durham University
- BBSRC Response Mode Grant Funding Panel Member
The role of the actin cytoskeleton in membrane trafficking and cell organization
Cells are the basic unit of life and all organisms are composed of one or more cells. Central to the functioning of many cells, including human cells, is the internal skeleton, or cytoskeleton. This cytoskeleton is required for cells to have certain shapes that are often a necessary part of their functioning. However, unlike our own body skeleton that is static, the cytoskeleton is able to remodel itself to change cell shape, or allow a cell to move.
One of the most important proteins in the cytoskeleton is called actin. It is an amazing protein because it is almost the same now as hundreds of millions of years ago, long before humans, or even vertebrates existed. Staying so similar over time is called evolutionary conservation. Proteins that are very important to cell functioning are the most highly conserved. Actin is a protein that can join together with other actin proteins to form long lines or filaments. These filaments can be organised by other proteins to form large structures that are part of the cytoskeleton.
We are interested in how this protein is controlled in cells and in particular, we are trying to determine how the filaments can be started from single actin proteins in a process called nucleation. We are also aiming to understand how force can be generated by these filaments to help inward bending of the outer membrane of the cell to let parts of the membrane to be pinched off (internalised) This process, called endocytosis allows the surface composition of the cell to be effectively regulated.
- Iwona Smaczynska-de Rooij
- Ellen Allwood
- John Palmer
Postgraduate PhD students:
- Joe Tyler
- Harriet Knafler
Scientific Questions we are addressing:
- How are new actin filaments initiated at cell membranes?
- What mechanisms ensure that endocytosis is a unidirectional process?
- How do dynamin, amphiphysin and actin function together during membrane scission?
- What alternative mechanisms of endocytosis are present in yeast
- What is the importance of endocytosis for virulence of the pathogenic fungus Candida albicans?
Techniques we use:
- Molecular biology: PCR, cloning, in vitro mutagenesis
- Biochemistry: protein expression and purification, binding assays including microscale thermophoresis; immunoprecipitation and GST pull down; lipid and liposome binding, actin polymerization assays, negative staining and electron microscopy analysis of actin filaments
- Genetics: S. cerevisiae and Candida albicans gene deletion and epitope tagging, growth, phenotypic analysis, strain crossing, tetrad dissection
- Cell Biology and microscopy: dual colour live cell imaging including quantitative analysis and patch tracking using Deltavision microscope; yeast endocytosis assays and vital staining for mitochondria and vacuoles.
- Electron microscopy, mostly in collaboration with Martin Goldberg, Durham
- BBRSC Project Grant, (2016 –2019) £638,000. Elucidating the molecular mechanism of Arp2/3-independent actin nucleation by WASP family proteins. Principle investigator.
- BBSRC Infrastructure Award (2013) A Quantitative Biomolecular Interaction Suite. Co-applicant.
- MRC Imaging Initiative (Sheffield IMAging) (2013) Co-applicant
- BBSRC Project Grant (2013 – 2016) Elucidating the mechanism of endocytic invagination and scission. Principle investigator
- BBSRC Project Grant (2012 – 2015) £602,000. Defining factors that ensure unidirectionality of endocytosis. Principle investigator
- Yorkshire Cancer Research (2010-2011) –Regulation of podosome/invadopodia dynamics in prostate cancer cells. Co-applicant.
- BBSRC Project Grant. (2009 - 2012). Endocytic Invagination and Vesicle Scission - interplay between Dynamin homologues and Amphiphysins in Budding Yeast. Principle investigator.
- MRC Senior non-clinical Fellowship Renewal (2007-2012) - £1,142,297. The Role of Actin in Cell Homeostasis.