Bacillus subtilis SepF Binds to the C-Terminus of FtsZ

Ewa Krol*, Sebastiaan P. van Kessel, Laura S. van Bezouwen, Neeraj Kumar, Egbert J. Boekema, Dirk-Jan Scheffers

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

36 Citations (Scopus)
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Abstract

Bacterial cell division is mediated by a multi-protein machine known as the "divisome", which assembles at the site of cell division. Formation of the divisome starts with the polymerization of the tubulin-like protein FtsZ into a ring, the Z-ring. Z-ring formation is under tight control to ensure bacteria divide at the right time and place. Several proteins bind to the Z-ring to mediate its membrane association and persistence throughout the division process. A conserved stretch of amino acids at the C-terminus of FtsZ appears to be involved in many interactions with other proteins. Here, we describe a novel pull-down assay to look for binding partners of the FtsZ C-terminus, using a HaloTag affinity tag fused to the C-terminal 69 amino acids of B. subtilis FtsZ. Using lysates of Escherichia coli overexpressing several B. subtilis cell division proteins as prey we show that the FtsZ C-terminus specifically pulls down SepF, but not EzrA or MinC, and that the interaction depends on a conserved 16 amino acid stretch at the extreme C-terminus. In a reverse pull-down SepF binds to full-length FtsZ but not to a FtsZ Delta C16 truncate or FtsZ with a mutation of a conserved proline in the C-terminus. We show that the FtsZ C-terminus is required for the formation of tubules from FtsZ polymers by SepF rings. An alanine-scan of the conserved 16 amino acid stretch shows that many mutations affect SepF binding. Combined with the observation that SepF also interacts with the C-terminus of E. coli FtsZ, which is not an in vivo binding partner, we propose that the secondary and tertiary structure of the FtsZ C-terminus, rather than specific amino acids, are recognized by SepF.

Original languageEnglish
Article numbere43293
Number of pages9
JournalPLoS ONE
Volume7
Issue number8
DOIs
Publication statusPublished - 13-Aug-2012

Keywords

  • CELL-DIVISION PROTEIN
  • ESCHERICHIA-COLI FTSZ
  • CYTOSKELETAL PROTEIN
  • RING
  • ZIPA
  • MINC
  • SITE
  • POLYMERIZATION
  • PROTOFILAMENTS
  • LOCALIZATION

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