PT - JOURNAL ARTICLE AU - Nam, Onyou AU - Musial, Sabina AU - Demulder, Manon AU - McKenzie, Caroline AU - Dowle, Adam AU - Dowson, Matthew AU - Barrett, James AU - Blaza, James N. AU - Engel, Benjamin D. AU - Mackinder, Luke C. M. TI - A Protein Blueprint of the Diatom CO<sub>2</sub>-Fixing Organelle AID - 10.1101/2023.10.26.564148 DP - 2024 Jan 01 TA - bioRxiv PG - 2023.10.26.564148 4099 - http://biorxiv.org/content/early/2024/09/24/2023.10.26.564148.short 4100 - http://biorxiv.org/content/early/2024/09/24/2023.10.26.564148.full AB - Diatoms are central to the global carbon cycle. At the heart of diatom carbon fixation is an overlooked organelle called the pyrenoid, where concentrated CO2 is delivered to densely packed Rubisco. Diatom pyrenoids fix approximately one-fifth of global CO2, but virtually nothing is known about this organelle in diatoms. Using large-scale fluorescence protein tagging and affinity purification-mass spectrometry, we generate a high-confidence spatially-defined protein-protein interaction network for the diatom pyrenoid. Within our pyrenoid interaction network are 10 proteins with no known function. We show that six of these form a static shell encapsulating the Rubisco matrix of the pyrenoid, with the shell critical for pyrenoid structural integrity, shape, and function. Although not conserved at a sequence level, the diatom pyrenoid shares some architectural similarities to prokaryotic carboxysomes. Collectively, our results support the convergent evolution of pyrenoids across the two main plastid lineages and uncover a major structural and functional component of global CO2 fixation.Competing Interest StatementThe authors have declared no competing interest.