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. 2009 Feb 5;1(2):219–233. doi: 10.18632/aging.100020

Figure 4. WRN ATPase/helicase, but not exonuclease activity, is required to restore the slow growth phenotype of top3 in sgs1 top3 background.

Figure 4.

Panel A, WRN protein with conserved domains and positions of site-directed mutations. Panel B, Expression of WRN and WRN variants in transformed sgs1 top3 was induced at 2% gal concentration and cells were harvested after 6 h. Equal amount of total cell lysate was loaded on to 8-16% polyacrylamide SDS gels, followed by Western blotting using anti-WRN antibody. sgs1 top3 strain transformed with ATPase/helicase-dead (YEp195SpGAL-WRN K577M), exonuclease-dead (YEp195SpGAL-WRN E84A), RQC mutant (YEp195SpGAL-WRN K1016A), or polymorphic mutant (YEp195SpGAL-WRN R834C) was streaked on SC-Trp plates containing either 2% glu (Panel E) or 2% gal (Panel D). Plates were incubated at 30°C for 2 days and then photographed. Composition of the plates was as in Panel C.