Ribosome assembly and maturation are tightly regulated to ensure functional ribosomes and translational accuracy. In bacteria, factors like IF2 and IF3 facilitate subunit association, while RsfS, an anti-association factor, prevents ribosome assembly under nutrient-limited conditions, aiding stress adaptation. Ribosome maturation factors also coordinate rRNA folding and protein incorporation to form active subunits. Disruptions in maturation pathways may redirect translation factors to reactivate translation machinery during stress. Understanding these dynamics is crucial, as the repurposing of these factors is key to bacterial stress response, though the mechanisms remain unclear. This proposal aims to explore ribosome assembly mechanisms, focusing on factors aiding reassembly under stress, especially in maturation factor-deficient strains, and the role of α form-IF2 in subunit association. We will use MS analysis and cryo-EM to quantify and visualize these factors, including IF2, across different growth phases and during 70S formation.

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