Extended Data Fig. 5: Dri expression and association with the ribosome.
From: Structure of an archaeal ribosome reveals a divergent active site and hibernation factor
a, Recombinantly expressed and purified Dri proteins resolved on a 4–12% polyacrylamide gel stained with Coomassie brilliant blue. b, P. calidifontis lysate-based in vitro translation assay. After translation of the HiBiT peptide, luminescence was measured from the HiBiT-LgBiT complex. For the no mRNA and buffer controls, 3 µL of Dri buffer A was added to the reaction. Each point on the graph represents an individual measurement, and data and error bars represent the mean and standard deviation of the three measurements, respectively. The fold change from the buffer control is indicated on top of each bar. A two-sided, two-sample t-test was used to compare values (p-values from left to right: 6x10−5, 3x10−6, 7x10−8, 4x10−8, 2x10−7, 4x10−8, 2x10−4, 9x10−5, 7x10−8). c, Sucrose gradient fractionation of ribosomes from P. calidifontis cultures grown to the indicated optical densities. Data was overlayed based on the 50S peak and grey lines bracket the sample that was collected for mass spectrometry. d, Fold change, measured by mass spectrometry, in protein levels from P. calidifontis 70S ribosomes samples isolated from stationary and logarithmic phase cultures. P-values were calculated with ANOVA in PEAKS Studio. e, The difference in Dri and uL2 mRNA Ct values from total RNA extracted from cultures at logarithmic (OD600 0.07) or stationary (OD600 0.15) phase, measured by RT-qPCR. Each point on the graph represents an individual measurement, and data and error bars represent the mean and standard deviation of the three measurements, respectively. A two-sided, two-sample t-test was used to compare values (p-value=0.046). f, Sucrose gradient fractionation of M. acetivorans ribosomes. Grey lines bracket the sample that was collected for mass spectrometry.
