A genomic and structural study of FtsZ constriction in cell division

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DESCRIPTION (provided by applicant): FtsZ (filamentous temperature sensitive Z) is an essential protein for bacterial cell division. It assembles the Z-ring that pinches the cell in two. I am interested in two different aspects of FtsZ function. 1. FtsZ is the only protein necessary for constriction force generation in liposomes in vitro. FtsZ from foreign bacteria and some mutant FtsZ were also able to cause cell division in E. coli. However, some forms of FtsZ tested required the acquisition of unknown genomic mutations to suppress the defective FtsZ phenotype and allow division. The mutations in these suppressor strains are currently unknown, but are likely loss-of- function mutations in other pathways involved in cell division. This project aims to utilize high-throughput sequencing technology to resequence the entire genome of each of the 13 suppressor strains to map the suppressor mutation and identify further genes of interest in bacterial cell division. 2. The FtsZ protein has a C-terminal (Ct) tail region that lacks a lacks a well-defined structure as visualized through X-ray crystallography, indicating that it serves primarily as a spacer to link the FtsZ globular domain to the FtsA binding site and the membrane. However, some FtsZ mutants with small 20aa insertions in the Ct tail are incapable of functioning for cell division. This project aims to examine the role of the Ct tail function in vivo and in vitro. The Ct tail will be expressed and purified apart from the FtsZ globular domain for NMR structure determination. Varied forms of FtsZ will be expressed using alternative Ct tail sequences and length, and the ability of the protein to produce a constriction force in liposomes in vitro, and to function for cell division in vivo will be assessed. Public Health Relevance: FtsZ is a required protein for cell division in most bacteria. A detailed understanding of FtsZ structure and function would increase the possibility and efficacy of targeting the protein pharmacologically to inhibit bacterial division, thus creating potential new antibiotics.