Author ORCID Identifier

0009-0003-2171-3376

Date of Graduation

12-2025

Document Type

Thesis (MS)

Program Affiliation

Microbiology and Infectious Diseases

Degree Name

Masters of Science (MS)

Advisor/Committee Chair

William Margolin

Committee Member

Heidi Kaplan

Committee Member

Peter Christie

Committee Member

Travis Moore

Committee Member

Anna Konovalova

Abstract

Bacterial cytokinesis requires precise coordination between cytoplasmic and periplasmic components of the divisome to ensure that membrane constriction and chromosome segregation occur in synchrony. The actin-like protein FtsA plays a central role in this process by tethering FtsZ to the membrane and recruiting downstream division proteins. Despite its essential function, the mechanisms by which FtsA coordinates divisome activation and the recruitment of late-acting proteins remain incompletely defined. To address this, the role of FtsA in activating septal synthesis and recruiting the DNA translocase FtsK was examined using mutational analysis in Escherichia coli.

The FtsAM96E R153D variant, which has previously been shown to disrupt double-stranded (DS) filament formation, was unable to support cell division but its defect was suppressed by the superfission allele ftsL, which constitutively activates the FtsWI complex. These results demonstrate that the DS filament form of FtsA is normally required for activating FtsWI and septal peptidoglycan synthesis but not for recruiting proteins to the divisome such as FtsK and FtsQLB, which in turn recruit and activate FtsWI. To further investigate FtsA-mediated recruitment of FtsK, mutagenesis of the FtsK N-terminal cytoplasmic domain identified two critical loop regions, Y106–D108 and E188–L190, essential for cell division. Mutations in these regions failed to fully complement an ftsKts strain, despite stable protein expression, indicating that these residues represent functional “hotspots” required for FtsK activity.

Together, these findings establish FtsA as a central regulator that couples activation of the septal synthase FtsWI with recruitment of FtsK, thereby synchronizing septal synthesis and chromosome segregation. This work provides new mechanistic insight into how FtsA integrates key steps of bacterial cytokinesis through its dual roles in activation and recruitment.

Keywords

bacterial genetics, cell division, divisome assembly, FtsA, FtsK

Included in

Bacteriology Commons

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