Membrane binding controls the ATPase cycle and localization of MinD in Bacillus subtilis
Data files
May 05, 2026 version files 1.09 GB
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Figure6–source_data_1.zip
1.09 GB
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README.md
3.59 KB
Abstract
This dataset includes all analysis done for this paper regarding single-molecule localization microscopy (SMLM) and single-particle tracking (SPT). It contains two main folders, "Figure 6" and "Figure 7", which contain all relevant data for the corresponding figures in the article. The folders and their structures were choosen in a way that it can be directly used to analyse the data in the free software tool "SMTracker 2," which was used to create the data for Figures 6 and 7 as well as Tables 4 and 5. Furthermore, it contains the raw localization tables of all SMLM data as well as all tracking data from SPT.
Dataset DOI: 10.5061/dryad.95x69p91b
Description of the data and file structure
The dataset was generated to investigate the dynamics and membrane interactions of the cell division protein MinD in Bacillus subtilis using single-molecule localization microscopy (SMLM) and single-particle tracking (SPT). Halo-tagged MinD was expressed in wild-type, ΔminC, and ΔminJ genetic backgrounds to assess the contributions of MinC and MinJ to MinD mobility, membrane association, and spatial organization in vivo. Furthermore, ATPase mutants of MinD (G12V, K16A, and D40A) as well as a membrane binding mutant (I260E) were investigated and compared to the wild-type.
Time-lapse fluorescence imaging was performed at high temporal resolution to capture individual Halo-MinD trajectories in living cells. The resulting datasets were analyzed to extract diffusion coefficients, population distributions of distinct mobility states (fast diffusive, slow diffusive, and confined), and stationary localization parameters.
Files and variables
File: Figure6–source_data_1.zip
Description: Contains all SMLM data aquired to produce Figure 6, Figure 7, Table 4 and Table 5 of the publication.
Folder: Figure 6
Description: Contains 5 subfolders that contain all necessary data for SMLM analysis of the MinD wild-type protein as well as for the mutants G12V, K16A, D40A and I260E. Each of the subfolders contains another 5 subfolders:
- Folder "cell_meshes": Contains the cell outlines for all cells in the field of view, necessary to link link and limit tracking data in the "SMTracker 2.0" software. Outlines were created in the free software "Oufti".
- Folder "ElyraTable": Contains raw tables for all SMLM localizations for every Field of View. This data was used to create heatmaps of all recorded Halo-MinD localizations, but is not required for analysis in "SMTracker 2.0".
- Folder "phase": Contains phase (widefield) images of every field of view. Required for analysis in "SMTracker 2.0".
- Folder "raw": Empty folder, but presence of the folder is required for analysis in "SMTracker 2.0". Raw microscopy data is not included because the dataset would be in the terrabyte size range, which is why we provided the raw SMLM tables instead (ElyraTable folder), which can be analysed without the proprietary Zen Black software.
- Folder "trackmate": Contains all tracking data, created using Trackmate 6.0.1 in Fiji (ImageJ2) 1.53 g. Required for analysis in "SMTracker 2.0".
Folder: Figure 7
Description: Contains 3 subfolders that contain all necessary data for SMLM analysis of the Halo-MinD protein expressed in a wild-type background as well as in a delta MinC or a delta MinJ background. Each of the subfolders contains the same 5 subfolders as described in "Folder 6."
Code/software
Analysis of the data used in Figure 6 and 7 and Table 4 and 5 requires the "SMTracker 2.0" software (https://sourceforge.net/projects/singlemoleculetracker/).
Both "Folder 6" and "Folder 7" should just be openend using the SMTracker. To do so, start the SMTracker and change these required settings:
- File -> type of data -> trackmate
- Pixel size [nm] -> 97
- Time intervals [ms] -> 24
Next, press: "select folder" and open either of the two folders "Figure 6" or "Figure7."