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. 2021 May 7;8:643696. doi: 10.3389/fmolb.2021.643696

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Copyright © 2021 Landwehr, Milanov, Angebauer, Hong, Jüngert, Hiersemenzel, Siebler, Schmit, Öztürk, Dannenmaier, Drepper, Warscheid and Koch.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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YchF counteracts the anti-associative effect of IF3 overexpression. (A) Wild-type and ΔychF cells were co-transformed with plasmids pQE30infC and pREP4. After induction of infC expression, cells were harvested, lysed; and the supernatants after centrifugation were analyzed by immune-detection using α-His antibodies against His-tagged IF3 and α-YchF antibodies. (B) The supernatants of the cells in panel (A) were subjected to sucrose-gradient density centrifugation and ribosomal fractions were monitored photometrically at 256 nm. Representative traces of at least two independent experiments are shown. (C) Quantification of the ribosomal fractions of wild-type and ΔychF strains expressing pQE30infC as shown in panel (B) (n = 2). The area below the curves was quantified using ImageJ and set to 100%. Areas corresponding to the 30 S, 50 S, and 70 S populations were extrapolated to the baseline, individually quantified and their relative abundance is shown.