Live imaging of a hyperthermophilic archaeon reveals distinct roles for two ESCRT-III homologs in ensuring a robust and symmetric division

Authors: Andre Arashiro Pulschen, Delyan R Mutavchiev, Siân Culley, Kim Nadine Sebastian, Jacques Roubinet, Marc Roubinet, Gabriel Tarrason Risa, Marleen van Wolferen, Chantal Roubinet, Uwe Schmidt, Gautam Dey, Sonja-Verena Albers, Ricardo Henriques, Buzz Baum
Technologies: Sulfoscope
Paper published in Current Biology, January 2020
Publisher: Elsevier

The manuscript by Pulschen et al. explores the division mechanism of the hyperthermophilic archaeon Sulfolobus acidocaldarius, focusing on the roles of two ESCRT-III homologs, CdvB1 and CdvB2. Using various techniques such as immunofluorescence microscopy, flow cytometry, and western blotting, the researchers discovered that CdvB1 is involved in the early stages of division, while CdvB2 plays a role in the late stages. The absence of CdvB2 leads to asymmetric division and an increase in division failure events. The study provides new insights into the division mechanism of archaeal cells and the role of ESCRT-III homologs in cell division. Techniques used included immunofluorescence microscopy on LabTek chambered slides, flow cytometry analysis on BD LSR II and BD Fortessa, and western blotting with lysis, centrifugation, transfer, blocking, primary and secondary antibodies, and development using Li-COR Odyssey Infrared Imaging System. Quantification and statistical analysis were conducted using GraphPad Prism, with non-parametric Mann–Whitney tests. The researchers studied changes in DNA organization, division failure, division speeds, asymmetry measurements, and the impact of different treatments on cell division.