(
A) Genes with putative function were visualized in UMAP space. MSMEG_0317 (
Cashmore et al., 2017) and MSMEG_6276 (murT) were found in the cell-wall cluster, supporting their suggested roles as components of cell-wall synthesis. The transcription factor, WhiA (
Rustad et al., 2014), was found in the DNA-Cell Division-Translation Cluster, suggesting a role in cell-cycle regulation. (
B) MSMEG_0317 produces a distinct phenotype on knockdown, with Dendra-tagged MSMEG_0317 localizing to the cell wall and cell septa. (
C) Knockdown of whiA produces elongated, branching cells, with Dendra-tagged WhiA localized to the nucleoid. (
D) MSMEG_6276, a gene predicted by HHpred (
Hildebrand et al., 2009) to have strong homology to murT/gatD, causes cellular bulging and lysis on silencing.