Proteome-wide Capture of Co-translational Protein Dynamics in Bacillus subtilis Using TnDR, a Transposable Protein-Dynamics Reporter
Proteome-wide Capture of Co-translational Protein Dynamics in Bacillus subtilis Using TnDR, a Transposable Protein-Dynamics Reporter
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DOI：
10.1016/j.celrep.2020.108250
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发表时间：
2020-10-13
期刊：
影响因子：
8.8
通讯作者：
Chiba, Shinobu
中科院分区：
文献类型：
--
作者：
Fujiwara, Keigo;Katagi, Yutaro;Chiba, Shinobu
动态蛋白质成熟，如定位，折叠和复合物的形成，可以同时发生。新生多肽在多大程度上参与共翻译动力学以产生功能性蛋白质组的互补物？我们解决这个问题，使用蛋白质动力学报告（DR）模块，包括一个力敏感的逮捕序列（枯草芽孢杆菌MifM），其次是LacZ的框架。携带DR的工程化转座子TnDR被转座到B中。枯草杆菌染色体上的蛋白质，以产生蛋白质的N-末端区域和C-末端DR模块之间的翻译融合。通过寻找LacZ+集落，我们确定了数百种取消延长停滞的蛋白质，最有可能反映了它们协同启动成熟/定位过程的能力。案例研究确定了B。在翻译完成之前启动与伴侣分子组装的枯草杆菌蛋白质。这些结果表明，共翻译成熟是一个经常发生的事件在蛋白质生物合成。
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 cotranslational maturation is a frequently occurring event in protein biogenesis.