Overview

Electron cryo-tomography of protein transport systems

We employ different techniques in electron cryo-microscopy to study the fundamental process of protein transport across cell membranes. We investigate a number of research themes - mitochondrial protein import, the bacterial type IV pilus assembly machinery and the mechanism of filamentous bacteriophage infection and egress. These are important topics that can help us to understand the principles of protein transport, and how mechanisms may be compromised in disease states or exploited on bacterial and viral infection.

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Biography

During her undergraduate summer project, Vicki Gold became fascinated by the concept of protein translocation across membranes. She embarked on a career starting at the University of Bristol, where she was awarded her PhD for studies on the general protein secretory (Sec) pathway in bacteria. After award of an EMBO Long-Term fellowship, she moved to the Max-Planck Institute of Biophysics in Frankfurt, Germany to work on mitochondrial protein import. During this time, the technique of electron cryo-microscopy was undergoing a so-called “resolution revolution”. The ability to determine protein structures at near-atomic level detail sparked her interest in the powerful method. Vicki developed her own research themes using electron cryo-microscopy to investigate protein transport systems and returned to the UK in 2017 as a Senior Lecturer and PI. Vicki is based in the Living Systems Institute.

Research Team

Dr. Alexander Neuhaus (PDRA) – the bacterial type IV pilus machinery

Dr. Rebecca Conners (PDRA) – the mechanism of filamentous phage infection and egress

Dr. Mathew McLaren (electron microscopy Experimental Officer)

Dr. Kelly Sanders (wet laboratory technician)

Emma Buzzard – (SWBio DTP student) - Investigating mitochondrial complex I assembly as a factor for disease

Qualifications

2008: PhD Biochemistry, University of Bristol
2004: BSc Biological Sciences, University of Warwick

Career

2016-present: Senior Lecturer, Living Systems Institute, University of Exeter, UK
2011-2016: Postdoctoral Fellow, Department of Structural Biology, Max Planck Institute of Biophysics, Germany
2008-2011: Postdoctoral Research Associate, School of Biochemistry, University of Bristol, UK    
2004-2007: Postgraduate Research Assistant, School of Biochemistry, University of Bristol, UK     

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Research

Research interests

Mitochondrial protein import

Mitochondria are the primary cellular source of ATP and form an important bioenergetic and metabolic signalling hub. Only 1% of mitochondrial proteins are encoded on mitochondrial DNA and the remainder are imported from the cytosol. Protein translocases are therefore essential for correct protein targeting and localisation, and a growing number of human pathologies have been linked to mitochondria and the import apparatus. We work to understand how protein import into mitochondria works by electron cryo-tomography, single-particle analysis, fluorescence microscopy and protein biochemistry. For further detail see Gold, V. et al, Nat Commun (2014), Gold, V. et al, EMBO Rep (2017) & Gold, V. et al, Methods Mol Biol (2017).

The bacterial type IV pilus assembly machinery

Type IV pili are several micron long filaments that power bacterial motility and adhesion. Pili thus enable bacteria to move within and interact with our environment, and are key determinants of virulence. The machinery that assembles type IV pili is a massive multi-membrane spanning complex, which in some species has a dual function in DNA uptake. This ability enables rapid genetic adaptation and the development of resistance to antimicrobials. We are using electron cryo-microscopy to study pili and the machinery that assembles them at high-resolution detail. For further detail see Gold, V. et al, eLife (2015) & Gold, V. & Kudrayashev, M., Curr Opin Struct Biol (2016) and Daum, B. et al, Biol Chem (2018).

Filamentous phage infection and egress

In a world becoming increasingly dominated by antibiotic resistant bacteria, treatment strategies need to be reassessed. We are using electron cryo-microscopy to study the filamentous phage (Ff) encoded assembly system, with the ultimate aim of developing cell-targeted antimicrobials. We are working to reveal structural details of the virus-host interaction, and of the machinery that enables the virus to exit the cell and proliferate.

Equipment and facilities

We are currently equipped with brand new state-of-the-art laboratories for cryoEM sample preparation, imaging and data processing. We employ a cryo- and automation-capable 120kV microscope (FEI Tecnai Spirit) with CMOS camera for screening samples and generation of initial single particle EM and cryoET data sets. For higher resolution data collection we use our shared GW4 200kV cryo-microscope (FEI Talos Arctica) fitted with a K2 direct electron detector at the South West Regional Facility for CryoEM. We are also well equipped with standard wet laboratory equipment for molecular biology, biochemistry, protein expression and purification. 

Research networks

• Prof. Agnieszka Chacinska, IIMCB Warsaw, Poland
• Prof. Ian Collinson, University of Bristol, UK
• Prof. Beate Averhoff, University of Frankfurt, Germany
• GW4 South West Regional Facility for CryoEM

Research grants

• 2019 Wellcome Trust
  Microbes produce long, hair-like appendages called flagella and pili, which project from their cell surface. Both types of filament are multifunctional and act as a means of communication between individual microbes, and between microbes and the environment. Flagella and pili also enable microbes to swim, attach to and “walk” along surfaces, exchange genetic material and form biofilms. Examples include dental plaque or dangerous contamination on hospital catheters and implants. As these filaments also enable microbes to move, they help microbes to spread and infect the human body. Study of the dynamic behaviour of these filaments will help us understand their function and inform the design of new drugs. This is especially important in view of the growing number of infectious bacteria that develop antibiotic resistance. Through recent work we have determined the structures of flagella and pili at very high detail, where their atomic composition can be seen. In this project, we will now explore how these atomic structures determine large-scale motion. This will provide targets for the development of new drugs, which can disrupt the motion and spread of infectious microbes.
• 2018 BBSRC
  The bacterial type IV pilus machinery as a DNA translocator
• 2018 Wellcome Trust
  The filamentous phage assembly system as a therapeutic tool
• 2017 Wellcome Trust
  Electron Cryo-microscopy equipment grant

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External Engagement and Impact

Administrative responsibilities

Chair of the Steering committee for the South West Regional Facility for CryoEM, GW4 and University of Exeter                  

Awards

EMBO Long-Term Fellowship, held at the Max-Planck Institute of Biophysics, Frankfurt am Main, Germany (2011-2013)

Lead PI on eBic BAG (Block Allocation Group) for time at the  Electron Bio-Imaging Centre (eBIC) at Diamond

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Teaching

Modules

2023/24

• BIO3077 - Frontiers in Molecular Cell Biology
• BIOM547 - Frontiers in Molecular Cell Biology

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Supervision / Group

Postdoctoral researchers

Postgraduate researchers

Research Technicians

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