Supplementary Figure 3: GTP hydrolysis is essential for release of HflX from the 50S subunit.
From: HflX is a ribosome-splitting factor rescuing stalled ribosomes under stress conditions
(a) Formation of 70S ribosome by association of vacant 30S (0.25 µM) with 50S (0.25 µM) in the absence (brown right triangle), or presence of HflX–GTP (black square) and HflX–GMPPNP (olive green down triangle), followed by increase in Rayleigh light scattering with time in a stopped flow apparatus. (b) The effect of HflX–GTP (black square) and HflX–GMPPNP (olive green down triangle) on the formation of 70S initiation complex. The experimental setup and data fitting were same as in a, except that an mRNA programmed 30S pre–IC was used instead of vacant 30S. As a control, the association reaction was also run without HflX (brown right triangle).
