SAGA1 and SAGA2 localize the starch sheath to the pyrenoid in <i>Chlamydomonas reinhardtii</i>

Most algae enhance their CO₂ assimilation by concentrating CO₂ within the pyrenoid, a biomolecular condensate that contains the CO₂-fixing enzyme Rubisco. Many pyrenoids are surrounded by a starch sheath that is thought to slow the escape of CO₂ from the pyrenoid, but how the starch sheath is localized to the pyrenoid remains poorly understood. Here, in the leading model alga <i>Chlamydomonas reinhardtii</i>, we find that the protein SAGA2 is necessary for early pyrenoid starch sheath biogenesis and works redundantly with its homolog, SAGA1, to localize the starch sheath to the pyrenoid. SAGA2 and SAGA1 were enriched in different regions of the pyrenoid-starch sheath interface, suggesting that they play complementary roles. Both <i>saga2</i> and <i>saga1</i> mutants showed defects in starch sheath coverage early during pyrenoid formation that were improved at a later timepoint. Strikingly, a <i>saga1;saga2</i> double mutant did not have a starch sheath around the pyrenoid and showed decreased overall starch content. SAGA1 and SAGA2 starch-binding domains bound to starch, the starch mimic molecule β-cyclodextrin, and the starch precursor molecule maltoheptaose, suggesting a role for SAGA1 and SAGA2 in starch granule initiation. We propose a model where SAGA1 and SAGA2 each locally prime starch sheath initiation in a distinct region of the pyrenoid surface by enriching starch precursor molecules around the pyrenoid. These findings advance the understanding of algal starch sheath biogenesis and provide insights into the associations between biomolecular condensates and other cellular structures.