Membrane remodelling during cell division in the thermoacidiophilic archaeaon Sulfolobus acidocaldarius

Lead Research Organisation: MRC Laboratory of Molecular Biology
Department Name: Cell Biology

Abstract

At division, the cellular membrane undergoes rapid and large-scale changes in shape and lipid composition. While these changes have been characterised in a number of eukaryotes and bacteria, almost nothing is known about the role of the membrane in archaeal cell division. Archaeal lipids are chemically distinct and membranes play an especially critical, structural role for archaeal cells that lack a cell wall. This is the case for Sulfolobus acidocaldarius, a model thermoacidophilic archaeon that is currently the closest experimentally tractable relative of eukaryotes. While cell division in Sulfolobus is driven by the activity of ESCRTIII and Vps4 - homologues of the proteins catalysing abscission in many eukaryotes - the lipid membrane that ESCRTIII remodels in Sulfolobus has a unique chemistry and organisation. Distinct from bacterial or eukaryotic lipids, archaeal lipids can span the membrane to create an effective monolayer. Using a combination of high temperature live-cell imaging, lipidomics and structural studies, I aim to determine the properties of the archaeal lipid membrane that are necessary to support ESCRTIII function in archaeal cell division. To do so I will: 1) create new tools for visualising membrane dynamics in archaea using both commercially available and newly synthesised lipid probes in live cell microscopy; 2) identify the spatial and temporal control of lipids and membrane properties during cell division using a combination of lipidomics, proteomics and microscopy; 3) characterise how ESCRTIII interacts with the archaeal membrane using structural techniques and in silico modelling. Through understanding of how conserved proteins can act on such different membrane architectures we hope to shed some light on fundamental principles of cell division as well as the evolution of the comparatively more complex division machinery of eukaryotes.

Publications

10 25 50
 
Description Alice published a paper describing the use of fluorescent probes for living imaging archaea and Sulfolobales at 75°C
Exploitation Route We published the methods so that others in the archaeal community can use them
Sectors Education

Other

 
Description Alice's work is still underway. However, she has made a number of important findings that we hope to bring together into one or more papers in the coming year. In addition, she has done much to advance the technology we use to image and analyse archaeal cells as they grow and divide. Her achievements thus far include: i) improvements to our microscope set-up (a SoRa spinning disk). ii) the development of better propbes for imaging the Sulfolobus membrane iii) lipidomics across the cycle and in various perturbed conditions iv) the phenotypic analysis of mutants and small molecules that interfere with cell shape and division in Sulfolobus. v) the development of an image analysis pipeline to analyse live imaging data (cell segmentation, tracking and morphometry)
First Year Of Impact 2023
Sector Education
Impact Types Societal

 
Description Membrane remodelling during cell division in the thermoacidiophilic archaeaon Sulfolobus acidocaldarius
Amount € 141,000 (EUR)
Funding ID ALTF_1041-2021 
Organisation European Molecular Biology Organisation 
Sector Charity/Non Profit
Country Germany
Start 01/2022 
End 12/2023
 
Title Improved Sulfoscope by addition of SoRa and the development of new dyes for live imaging 
Description We improved the Sulfoscope for archaeal live cell imaging 
Type Of Material Technology assay or reagent 
Year Produced 2021 
Provided To Others? Yes  
Impact We have shared the method extensively with the community 
 
Title Microfluidic devices for live imaging of archaea over long periods. 
Description We have developed microfabricated CHIPs with Morgan Delarue for imaging archaea live. We have shown that dyes can be added to label cells. We have redesigned our heated cap so we can image cells in devices without fluorescence. 
Type Of Material Technology assay or reagent 
Year Produced 2024 
Provided To Others? No  
Impact This will allow longterm imaging of archaea 
 
Description Analysis of transmembrane proteins across the archaea 
Organisation Dutch Research Council
Department Royal Netherlands Institute for Sea Research
Country Netherlands 
Sector Academic/University 
PI Contribution We have been working with Josh Hamm (NIOZ) and Juan Pierella Karlusich (Harvard) to determine complete set of archaeal TM proteins
Collaborator Contribution We are advising.
Impact None yet...
Start Year 2022
 
Description Analysis of transmembrane proteins across the archaea 
Organisation Massachusetts Institute of Technology
Country United States 
Sector Academic/University 
PI Contribution We have been working with Josh Hamm (NIOZ) and Juan Pierella Karlusich (Harvard) to determine complete set of archaeal TM proteins
Collaborator Contribution We are advising.
Impact None yet...
Start Year 2022
 
Description Archaeal cell-cell interactions and their role in the emergence of eukaryotes 
Organisation Brandeis University
Country United States 
Sector Academic/University 
PI Contribution We have come together to use archaea as a model of eukaryogenesis
Collaborator Contribution We co-wrote a grant that I lead
Impact We have published a number of papers that have benefited from the work of the team. We have also had a number of lab exchanges through this newwork.
Start Year 2019
 
Description Archaeal cell-cell interactions and their role in the emergence of eukaryotes 
Organisation Dutch Research Council
Department Royal Netherlands Institute for Sea Research
Country Netherlands 
Sector Academic/University 
PI Contribution We have come together to use archaea as a model of eukaryogenesis
Collaborator Contribution We co-wrote a grant that I lead
Impact We have published a number of papers that have benefited from the work of the team. We have also had a number of lab exchanges through this newwork.
Start Year 2019
 
Description Gordon and Betty Moore Foundation grant to study symbiosis 
Organisation Dutch Research Council
Department Royal Netherlands Institute for Sea Research
Country Netherlands 
Sector Academic/University 
PI Contribution We were awarded a grant as part of the Symbiosis in Aquatic Systems Initiative.
Collaborator Contribution the collaboration is led by Anja Spang, it includes Dina Grohmann, Harald Huber and Laura Villanueva
Impact We are starting to share ideas and resources
Start Year 2020
 
Description Laura Villanueva 
Organisation Dutch Research Council
Department Royal Netherlands Institute for Sea Research
Country Netherlands 
Sector Academic/University 
PI Contribution We have a joint Simons Moore foundation grant together. We also hosted Kerstin Fiege from Laura's team at the LMB
Collaborator Contribution Laura's team are helping us with archaeal lipidomics.
Impact We have lipidomic data for Alice's work in the lab. Kerstin made progress with the cell biological of her symbiosis.
Start Year 2022
 
Description Lipid extraction from Sulfolbus with Sonja Albers 
Organisation Albert Ludwig University of Freiburg
Country Germany 
Sector Academic/University 
PI Contribution We have a joint VW grant with Sonja Albers.
Collaborator Contribution As part of this, Alice received lipids and a protocol from the Albers lab.
Impact We have yet to make use of the lipid fractions. We may have to repeat the extraction.
Start Year 2021
 
Description VW Life? 
Organisation Albert Ludwig University of Freiburg
Department Centre for Biological Signalling Studies (BIOSS)
Country Germany 
Sector Academic/University 
PI Contribution We were awarded a VW Life? award. Towards this we contributed to writing of the grant and preliminary data.
Collaborator Contribution the grant is led by Sonja-Verena Albers, and the team includes Jan Löwe, Lars-Oliver Essen and Andela Saric
Impact We are due for our last meeting in Austria this May. This will be a chance to review the ongoing work we are doing together. We expect to continue collaborating with Sonja Albers, Andela Saric and Jan Lowe.
Start Year 2019
 
Description VW Life? 
Organisation Medical Research Council (MRC)
Department MRC Laboratory of Molecular Biology (LMB)
Country United Kingdom 
Sector Academic/University 
PI Contribution We were awarded a VW Life? award. Towards this we contributed to writing of the grant and preliminary data.
Collaborator Contribution the grant is led by Sonja-Verena Albers, and the team includes Jan Löwe, Lars-Oliver Essen and Andela Saric
Impact We are due for our last meeting in Austria this May. This will be a chance to review the ongoing work we are doing together. We expect to continue collaborating with Sonja Albers, Andela Saric and Jan Lowe.
Start Year 2019
 
Description VW Life? 
Organisation Philipp University of Marburg
Country Germany 
Sector Academic/University 
PI Contribution We were awarded a VW Life? award. Towards this we contributed to writing of the grant and preliminary data.
Collaborator Contribution the grant is led by Sonja-Verena Albers, and the team includes Jan Löwe, Lars-Oliver Essen and Andela Saric
Impact We are due for our last meeting in Austria this May. This will be a chance to review the ongoing work we are doing together. We expect to continue collaborating with Sonja Albers, Andela Saric and Jan Lowe.
Start Year 2019
 
Description Visited NIOZ and collected samples in the field 
Organisation Dutch Research Council
Department Royal Netherlands Institute for Sea Research
Country Netherlands 
Sector Academic/University 
PI Contribution Alice Cezanne and I came to NIOZ for a joint meeting. We spent a day collecting anaerobic archaeal samples from a site near the sea with Josh Hamm
Collaborator Contribution Alice Cezanne and I came to NIOZ for a joint meeting. We spent a day collecting anaerobic archaeal samples from a site near the sea with Josh Hamm
Impact There are some cells in a flask at NIOZ. We hope they will grow up. We are also planning the next visit of Florian Mayer to the LMB in Cambridge.
Start Year 2023
 
Description Alice presented her work to School Children from Cambridgeshire 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Alice presented a picture of life in the lab to local school kids as part of the LMB enegagement activities
Year(s) Of Engagement Activity 2022,2023,2024
 
Description Archaea.bio 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Alice has been instrumental in setting up a website for the archaeal community which enables sharing of data and ESCPECIALLY of protocols.
Year(s) Of Engagement Activity 2023,2024
 
Description GRC archaeal meeting - talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Alice presented her latest data to the community
Year(s) Of Engagement Activity 2023
 
Description Talk MPI Tubingen 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I was invited to visit the MPI in Tubingen as a guest speaker.
Year(s) Of Engagement Activity 2023
 
Description Talk Munich 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I was an invited speaker at LMU Munich
Year(s) Of Engagement Activity 2023
 
Description Talk at Basel Biozentrum 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I was an invited speaker at Biozentrum
Year(s) Of Engagement Activity 2023