FIG 1.
Set1 and Set2 interact with RNA in vivo. (A) Domain organization of fusion proteins used in this study. RRM, RNA recognition motif; H4i, H4-interacting domain; AWS, associated with SET; PS, post-SET; WW, tryptophan-rich domain; CC, coiled-coil domain; SRI, Set2-Rpb1 interaction domain; PTH, protein A-TEV-His6 tag; HTP, His6-TEV-protein A tag. PSET1 and PSET2 are SET1 and SET2 promoters, respectively. (B) Western blots showing the protein abundance in the samples used in panel C. Cells were grown in minimal medium lacking tryptophan and UV cross-linked. The input lysate was analyzed with antibodies against H3K4me3, H3, and Pgk1 (loading controls). Molecules eluted from IgG beads using TEV protease were analyzed with anti-Set1 antibodies. (C and D) SDS-PAGE and autoradiography of the 5′-32P-labeled, cross-linked RNAs after purification of the tagged proteins or after mock purification of proteins from the untagged strain (BY4741).