PT - JOURNAL ARTICLE AU - A. Santana-Sánchez AU - L. Nikkanen AU - G. Toth AU - M. Ermakova AU - S. Kosourov AU - J. Walter AU - M. He AU - E-M. Aro AU - Y. Allahverdiyeva TI - Flv3A facilitates O<sub>2</sub> photoreduction and affects H<sub>2</sub> photoproduction independently of Flv1A in diazotrophic <em>Anabaena</em> filaments AID - 10.1101/2021.12.15.472848 DP - 2021 Jan 01 TA - bioRxiv PG - 2021.12.15.472848 4099 - http://biorxiv.org/content/early/2021/12/16/2021.12.15.472848.short 4100 - http://biorxiv.org/content/early/2021/12/16/2021.12.15.472848.full AB - The model heterocyst-forming filamentous cyanobacterium, Anabaena sp. PCC 7120 (Anabaena) represents multicellular organisms capable of simultaneously performing oxygenic photosynthesis in vegetative cells and the O2-sensitive N2-fixation inside the heterocysts. The flavodiiron proteins (FDPs) have been shown to participate in photoprotection of photosynthesis by driving excess electrons to O2 (Mehler-like reaction). Here, we addressed the physiological relevance of the vegetative cell-specific Flv1A and Flv3A on the bioenergetic processes occurring in diazotrophic Anabaena under variable CO2. We demonstrate that both Flv1A and Flv3A are required for proper induction of the Mehler-like reaction upon a sudden increase in light intensity, which is likely important for the activation of carbon-concentrating mechanisms (CCM) and CO2 fixation. Under low CO2 diazotrophic conditions, Flv3A is capable of mediating moderate O2 photoreduction, independently of Flv1A, but in coordination with Flv2 and Flv4. Strikingly, the lack of Flv3A resulted in strong downregulation of the heterocyst-specific uptake hydrogenase, which led to enhanced H2 photoproduction under both oxic and micro-oxic conditions. These results reveal a novel regulatory network between the Mehler-like reaction and the H2 metabolism, which is of great interest for future photobiological production of H2 in Anabaena.