Figure 2: Drg1-21 is unable to perform its function in pre-60S particle maturation. | Scientific Reports

Figure 2: Drg1-21 is unable to perform its function in pre-60S particle maturation.

From: A conserved inter-domain communication mechanism regulates the ATPase activity of the AAA-protein Drg1

Figure 2

(a) Drg1-21 shows reduced binding to pre-60s particles. Tandem affinity purifications of pre-60S particles using Arx1-TAP as bait protein were performed from strains expressing drg1-21, drg1-18 or wildtype DRG1 from their endogenous promoter. The protein composition of the purified particles was analysed by SDS-PAGE and western blotting with antibodies directed against the indicated factors. (b) Drg1-21 overexpression causes accumulation of shuttling proteins and export factors on pre-60S particles. Tandem affinity purifications of pre-ribosomal Arx1-TAP particles were performed from DRG1 wildtype strains containing plasmids for overexpression (GST-Drg1 or GST-Drg1-21) or the empty vector control. Cells were grown to an OD600 of 1.0 (early log-phase) and overexpression of the GST-Drg1 variants from the CUP1 promoter was induced by the addition of 0.5 mM CuSO4 and incubation for 3 hours. Original full-length images of cropped blots are provided in Supplementary Figs S7 and S8.

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