4.7 Article

The SpoVA membrane complex is required for dipicolinic acid import during sporulation and export during germination

Journal

GENES & DEVELOPMENT
Volume 36, Issue 9-10, Pages 634-646

Publisher

COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1101/gad.349488.122

Keywords

sporulation; germination; dormancy; dipicolinic acid; DPA; spoVA

Funding

  1. DARPA [F32GM130003, GM086466]
  2. Harvard Medical School Dean's Initiative [GM127399]
  3. National Institutes of Health
  4. [HR001117S0029]

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Researchers investigated the minimal set of proteins encoded in the Bacillus subtilis spoVA operon required for small molecule dipicolinic acid (DPA) import, which plays a central role in stress resistance and germination of dormant spores. They found that these proteins form a membrane complex, and proposed a model where DPA transport into spores involves cycles of unplugging and replugging the membrane channel, triggered by nutrient detection during germination to release DPA.
In response to starvation, endospore-forming bacteria differentiate into stress-resistant spores that can remain dormant for years yet rapidly germinate and resume growth in response to nutrients. The small molecule dipicolinic acid (DPA) plays a central role in both the stress resistance of the dormant spore and its exit from dormancy during germination. The spoVA locus is required for DPA import during sporulation and has been implicated in its export during germination, but the molecular bases are unclear. Here, we define the minimal set of proteins encoded in the Bacillus subtilis spoVA operon required for DPA import and demonstrate that these proteins form a membrane complex. Structural modeling of these components combined with mutagenesis and in vivo analysis reveal that the C and Eb subunits form a membrane channel, while the D subunit functions as a cytoplasmic plug. We show that point mutations that impair the interactions between D and the C-Eb membrane complex reduce the efficiency of DPA import during sporulation and reciprocally accelerate DPA release during germination. Our data support a model in which DPA transport into spores involves cycles of unplugging and then replugging the C-Eb membrane channel, while nutrient detection during germination triggers DPA release by unplugging it. Here, Gao et al. investigated the minimal set of proteins encoded in the Bacillus subtilis spoVA operon required for small molecule dipicolinic acid (DPA) import, which plays a central role in both the stress resistance of the dormant spore and its exit from dormancy during germination, and demonstrate that these proteins form a membrane complex. The authors propose a model in which DPA transport into spores involves cycles of unplugging then replugging the C-Eb membrane channel, while nutrient detection during germination triggers DPA release by unplugging it.

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