By the same authors

From the same journal

From the same journal

The Structure and Interactions of SpoIISA and SpoIISB, a Toxin-Antitoxin System in Bacillus subtilis

Research output: Contribution to journalArticle

Author(s)

Department/unit(s)

Publication details

JournalJournal of Biological Chemistry
DatePublished - 25 Feb 2011
Issue number8
Volume286
Number of pages12
Pages (from-to)6808-6819
Original languageEnglish

Abstract

Spore formation in Bacillus subtilis begins with an asymmetric cell division, following which differential gene expression is established by alternative compartment-specific RNA polymerase sigma factors. The spoIISAB operon of B. subtilis was identified as a locus whose mutation leads to increased activity of the first sporulation-specific sigma factor, sigma(F). Inappropriate spoIISA expression causes lysis of vegetatively growing B. subtilis cells and Escherichia coli cells when expressed heterologously, effects that are countered by co-expression of spoIISB, identifying SpoIISA-SpoIISB as a toxin-antitoxin system. SpoIISA has three putative membrane-spanning segments and a cytoplasmic domain. Here, the crystal structure of a cytoplasmic fragment of SpoIISA (CSpoIISA) in complex with SpoIISB has been determined by selenomethionine-multiwavelength anomalous dispersion phasing to 2.5 angstrom spacing, revealing a CSpoIISA(2) . SpoIISB(2) heterotetramer. CSpoIISA has a single domain alpha/beta structure resembling a GAF domain with an extended alpha-helix at its N terminus. The two CSpoIISA protomers form extensive interactions through an intermolecular four-helix bundle. Each SpoIISB chain is highly extended and lacking tertiary structure. The SpoIISB chains wrap around the CSpoIISA dimer, forming extensive interactions with both CSpoIISA protomers. CD spectroscopy experiments indicate that SpoIISB is a natively disordered protein that adopts structure only in the presence of CSpoIISA, whereas surface plasmon resonance experiments revealed that the CSpoIISA . SpoIISB complex is stable with a dissociation constant in the nanomolar range. The results are interpreted in relation to sequence conservation and mutational data, and possible mechanisms of cell killing by SpoIISA are discussed.

    Research areas

  • GRAM-POSITIVE BACTERIA, ESCHERICHIA-COLI, CRYSTAL-STRUCTURE, MOLECULAR-GRAPHICS, STATIONARY-PHASE, DIFFRACTION DATA, GAF DOMAIN, EXPRESSION, COMPLEX, PROTEIN

Discover related content

Find related publications, people, projects, datasets and more using interactive charts.

View graph of relations