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

• Published in: Genes & development 2022-05, Vol.36 (9-10), p.634-646
• Main Authors: Gao, Yongqiang, Barajas-Ornelas, Rocio Del Carmen, Amon, Jeremy D, Ramírez-Guadiana, Fernando H, Alon, Assaf, Brock, Kelly P, Marks, Debora S, Kruse, Andrew C, Rudner, David Z
• Format: Article
• Language: English
• Subjects: Bacillus subtilis - genetics; Bacillus subtilis - metabolism; Bacterial Proteins - metabolism; Picolinic Acids - metabolism; Research Paper; Spores, Bacterial - genetics
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/GAD.349488.122

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 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 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.