MinC N- and C-Domain Interactions Modulate FtsZ Assembly, Division Site Selection, and MinD-Dependent Oscillation in Escherichia coli

• Published in: Journal of bacteriology 2019-02, Vol.201 (4)
• Main Authors: LaBreck, Christopher J, Conti, Joseph, Viola, Marissa G, Camberg, Jodi L
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphatases - metabolism; Amino acids; Assembly; Bacteria; Bacterial Proteins - metabolism; Bacteriology; Cell Division; Coliforms; Complex formation; Cytoskeletal Proteins - metabolism; DNA Mutational Analysis; Domains; E coli; Escherichia coli; Escherichia coli - growth & development; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fluorescence; Genes, Reporter; Green fluorescent protein; Green Fluorescent Proteins - analysis; Green Fluorescent Proteins - genetics; Guanosine triphosphate; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mutagenesis; Mutation; Phospholipids; Polymerization; Polymers; Protein Binding; Protein Interaction Mapping; Proteins; Recombinant Fusion Proteins - analysis; Recombinant Fusion Proteins - genetics; Site selection
• ISSN: 0021-9193; 1098-5530
• DOI: 10.1128/JB.00374-18

The Min system in , consisting of MinC, MinD, and MinE proteins, regulates division site selection by preventing assembly of the FtsZ-ring (Z-ring) and exhibits polar oscillation MinC antagonizes FtsZ polymerization, and , the cellular location of MinC is controlled by a direct association with MinD at the membrane. To further understand the interactions of MinC with FtsZ and MinD, we performed a mutagenesis screen to identify substitutions in that are associated with defects in cell division. We identified amino acids in both the N- and C-domains of MinC that are important for direct interactions with FtsZ and MinD , as well as mutations that modify the observed oscillation of green fluorescent protein (GFP)-MinC. Our results indicate that there are two distinct surface-exposed sites on MinC that are important for direct interactions with FtsZ, one at a cleft on the surface of the N-domain and a second on the C-domain that is adjacent to the MinD interaction site. Mutation of either of these sites leads to slower oscillation of GFP-MinC , although the MinC mutant proteins are still capable of a direct interaction with MinD in phospholipid recruitment assays. Furthermore, we demonstrate that interactions between FtsZ and both sites of MinC identified here are important for assembly of FtsZ-MinC-MinD complexes and that the conserved C-terminal end of FtsZ is not required for MinC-MinD complex formation with GTP-dependent FtsZ polymers. Bacterial cell division proceeds through the coordinated assembly of the FtsZ-ring, or Z-ring, at the site of division. Assembly of the Z-ring requires polymerization of FtsZ, which is regulated by several proteins in the cell. In , the Min system, which contains MinC, MinD, and MinE proteins, exhibits polar oscillation and inhibits the assembly of FtsZ at nonseptal locations. Here, we identify regions on the surface of MinC that are important for contacting FtsZ and destabilizing FtsZ polymers.