Neuroscience Correction for “A yeast functional screen predicts new candidate ALS disease genes,” by Julien Couthouis, Michael P. Hart, James Shorter, Mariely DeJesus-Hernandez, Renske Erion, Rachel Oristano, Annie X. Liu, Daniel Ramos, Niti Jethava, Divya Hosangadi, James Epstein, Ashley Chiang, Zamia Diaz, Tadashi Nakaya, Fadia Ibrahim, Hyung-Jun Kim, Jennifer A. Solski, Kelly L. Williams, Jelena Mojsilovic-Petrovic, Caroline Ingre, Kevin Boylan, Neill R. Graff-Radford, Dennis W. Dickson, Dana Clay-Falcone, Lauren Elman, Leo McCluskey, Robert Greene, Robert G. Kalb, Virginia M.-Y. Lee, John Q. Trojanowski, Albert Ludolph, Wim Robberecht, Peter M. Andersen, Garth A. Nicholson, Ian P. Blair, Oliver D. King, Nancy M. Bonini, Vivianna Van Deerlin, Rosa Rademakers, Zissimos Mourelatos, and Aaron D. Gitler, which was first published November 7, 2011; 10.1073/pnas.1109434108 (Proc. Natl. Acad. Sci. U.S.A. 108, 20881–20890).
The authors note that Fig. 4D appeared incorrectly: “The experiments in this figure provide evidence that the RNA-binding protein TAF15 is aggregation-prone in vitro similar to RNA-binding proteins TDP-43 and FUS. When assembling the figure, in panel D, the 30-min TDP-43 image was taken from a 10-min image by mistake. We went back to the original EM images and have corrected Fig. 4D with the correct 30-min image.”
Fig. 4.
TAF15 is an aggregation-prone protein like TDP-43 and FUS. (A) Following TEV protease cleavage to remove the N-terminal GST tag, FUS, TAF15, and TDP-43 proteins were processed for SDS-PAGE and Coomassie stained to confirm purity and expected molecular weight. (B) GST-TDP-43, GST-FUS, or GST-TAF15 (3 μM) were incubated in the presence or absence of TEV protease at 25 °C for 0 to 90 min with agitation. Note that very little aggregation occurs in the absence of TEV protease. The extent of aggregation was determined by turbidity. Values represent means ± SEM (n = 3). (C) GST-TDP-43, GST-FUS, or GST-TAF15 (3 μM) was incubated in the presence of TEV protease at 25 °C for 0 to 60 min. At the indicated times, reactions were processed for sedimentation analysis. Pellet and supernatant fractions were resolved by SDS-PAGE and stained with Coomassie Brilliant Blue. The amount of protein in the pellet fraction was determined by densitometry in comparison with known quantities of the appropriate protein. Values represent means ± SEM (n = 3). A human RRM protein, DND1, which did not aggregate and was not toxic in yeast (Fig. 1 C and D), was also soluble and did not form aggregates in this assay. (D) GST-TDP-43, GST-FUS, or GST-TAF15 (3 μM) was incubated in the presence of TEV protease at 25 °C for 0 to 60 min. At various times, reactions were processed for EM. Small arrows denote small pore-shaped oligomers, and large arrows denote linear polymers. (Scale bar, 500 nm.) (E) Gallery of TDP-43, FUS, and TAF15 oligomers formed during aggregation reactions. (Scale bar, 50 nm.) (F) Following TEV protease cleavage to remove the N-terminal GST tag, TAF15 wild-type (WT), G391E, and R408C proteins were processed for SDS-PAGE and Coomassie stained to confirm purity and expected molecular weight. (G) ALS-linked TAF15 variants G391E and R408C displayed accelerated aggregation kinetics, whereas the R388H variant found in both cases and controls aggregated with similar kinetics to WT TAF15.
The corrected Fig. 4 and its legend appear below. The online version has been corrected.