Genome-wide screening reveals a novel class of carbonic anhydrase-like inorganic carbon transporters in chemoautotrophic bacteria

Many bacterial autotrophs rely on CO2 concentrating mechanisms (CCMs) to assimilate carbon. Although many CCM proteins have been identified, including a 200+ MDa protein organelle called the carboxysome, a systematic screen of CCM components has not been carried out. Here, we performed a genome-wide barcoded transposon screen to identify essential and CCM-related genes in the gamma-proteobacterium H. neapolitanus. Our screen revealed an operon encoding a domain of unknown function (PFAM:PF10070) and putative cation transporter subunit (PFAM:PF00361) is critical for CCM function. These two proteins, which we name DabA and DabB for "DABs accumulate bicarbonate," function as a heterodimeric, energy-coupled inorganic carbon pump in E. coli. Furthermore, DabA has distant homology to a beta-carbonic anhydrase and binds a zinc ion necessary for activity. Based on these results, we propose that DABs function as vectorial CAs coupled to cation gradients and serve as inorganic carbon pumps throughout prokaryotic phyla.