Figure S6.
The C Terminus of Ela1 Contains a UbH Domain, Specific to the CUE Domain of Def1, Related to Figure 6
(A) Western blot to detect binding to immobilized, recombinant Dsk2 and Def11-500, and ubiquitin-binding domains from Rap80 and Ataxin-3, in chromatin-enriched extracts from yeast cells expressing Myc-tagged Elc1. Membrane probed for Ela1 (anti-Ela1) and Elc1 (anti-myc) (upper two panels), as well as ubiquitylated proteins (anti-ubiquitin) (lower panel). Def1 pulls Ela1-Elc1 out of the extract, but does not bind significant amounts of ubiquitylated proteins, while Dsk2, Rap80, and Ataxin-3 pull unknown ubiquitylated proteins out of the extract, but do not bind significant amounts of Ela1-Elc1.
(B) Alignment of Ela1’s C-terminal region with yeast ubiquitin (alignment performed using ClustalW2) (Larkin et al., 2007).
(C) Western blot detecting binding of Myc-Elc1 to WT Ela1 (lanes 1-3) and Ela11-250 (lanes 4-6) in vivo. Note that while Elc1 binding to the shorter form of Ela1 is normal, binding to Rpb1 is appreciably decreased in ela11-250 cells.