Spatiotemporal localization of proteins in mycobacteria

• Published in: Cell reports (Cambridge) 2021-12, Vol.37 (13), p.110154-110154, Article 110154
• Main Authors: Zhu, Junhao, Wolf, Ian D., Dulberger, Charles L., Won, Harim I., Kester, Jemila C., Judd, Julius A., Wirth, Samantha E., Clark, Ryan R., Li, Yawei, Luo, Yuan, Gray, Todd A., Wade, Joseph T., Derbyshire, Keith M., Fortune, Sarah M., Rubin, Eric J.
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - metabolism; Cell Cycle; fluorescent protein; matrix factorization; microscopy image analysis; Molecular Imaging - methods; mycobacteria; Mycobacterium smegmatis; Mycobacterium smegmatis - metabolism; Organelles - metabolism; Protein Transport; Spatio-Temporal Analysis
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2021.110154

Although prokaryotic organisms lack traditional organelles, they must still organize cellular structures in space and time, challenges that different species solve differently. To systematically define the subcellular architecture of mycobacteria, we perform high-throughput imaging of a library of fluorescently tagged proteins expressed in Mycobacterium smegmatis and develop a customized computational pipeline, MOMIA and GEMATRIA, to analyze these data. Our results establish a spatial organization network of over 700 conserved mycobacterial proteins and reveal a coherent localization pattern for many proteins of known function, including those in translation, energy metabolism, cell growth and division, as well as proteins of unknown function. Furthermore, our pipeline exploits morphologic proxies to enable a pseudo-temporal approximation of protein localization and identifies previously uncharacterized cell-cycle-dependent dynamics of essential mycobacterial proteins. Collectively, these data provide a systems perspective on the subcellular organization of mycobacteria and provide tools for the analysis of bacteria with non-standard growth characteristics. [Display omitted] •MOMIA and GEMATRIA efficiently model mycobacterial protein localization•Polar exclusion of mycobacterial ribosomes relies on active translation•GEMATRIA reveals spatial partitioning of mycobacterial membrane proteins Zhu et al. develop a two-stage image analysis pipeline, MOMIA and GEMATRIA, that efficiently models the spatial and temporal dynamics of over 700 conserved proteins in M. smegmatis. Through the analysis they report spatial constraints of mycobacterial ribosomes and membrane complexes and reconstruct temporal dynamics from still image data.