Spatiotemporal localization of proteins in mycobacteria

Junhao Zhu, Ian D. Wolf, Charles L. Dulberger, Harim I. Won, Jemila C. Kester, Julius A. Judd, Samantha E. Wirth, Ryan R. Clark, Yawei Li, Yuan Luo, Todd A. Gray, Joseph T. Wade, Keith M. Derbyshire, Sarah M. Fortune*, Eric J. Rubin

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


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.

Original languageEnglish (US)
Article number110154
JournalCell reports
Issue number13
StatePublished - Dec 28 2021


  • Mycobacterium smegmatis
  • fluorescent protein
  • matrix factorization
  • microscopy image analysis
  • mycobacteria

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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