1. Functional mutants of ABCE1 are potent ribosome splitting factors.
Functional mutants of ABCE1 are potent ribosome splitting factors. 70S splitting efficiency of ABCE1 with functional mutations of the catalytic base was assayed by SDG and quantified by the peak height ratios. In the presence of AMP-PNP (pink trace), both variants ABCE1E238A (A) and ABCE1E485A (B) are significantly more potent ribosome splitting factors than WT ABCE1 (compare Fig 3C). Major difference between site I and site II is revealed in samples with ADP (green trace). Here, only ABCE1E485A with site II in an occluded state is able to split 70S, whereas ABCE1E238A has a significantly lower activity. Both factors require the presence of excess release factor showing that splitting is an active and specific process (>4% splitting in the absence of aRF1, blue trace). Representative set of three independent experiments.
Elina Nürenberg-Goloub et al. LSA 2018;1:e
© 2018 Nürenberg-Goloub et al.