Proteome-wide Capture of Co-translational Protein Dynamics in Bacillus subtilis Using TnDR, a Transposable Protein-Dynamics Reporter

Dynamic protein maturation, such as localization, folding, and complex formation, can occur co-translationally. To what extent do nascent polypeptides engage in the co-translational dynamics to produce the functional proteome’s complement? We address this question using a protein-dynamics reporter (DR) module comprising a force-sensitive arrest sequence (Bacillus subtilis MifM) followed in frame by LacZ. An engineered transposon, TnDR, carrying DR, is transposed into the B. subtilis chromosome to create translational fusions between N-terminal regions of proteins and the C-terminal DR module. By looking for LacZ+ colonies, we identify hundreds of proteins that cancel the elongation arrest, most probably reflecting their ability to initiate the maturation/localization process co-translationally. Case studies identify B. subtilis proteins that initiate assembly with a partner molecule before completion of translation. These results suggest that co-translational maturation is a frequently occurring event in protein biogenesis. [Display omitted] •TnDR is an arrest-sequence-based and transposable protein-dynamics reporter•TnDR captures co-translational protein localization and assembly•Proteome-wide screening has been conducted•Co-translational protein maturation is a frequently occurring event in the cell Fujiwara et al. combine a transposon and a force-sensitive arrest sequence to develop TnDR, a transposable protein-dynamics reporter. Proteome-wide screening identifies hundreds of B. subtilis proteins that co-translationally engage in a dynamic process of maturation (localization/assembly). They show that the co-translational occurrence is frequent for protein maturation in cells.