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. 2006 Jan;26(2):523–534. doi: 10.1128/MCB.26.2.523-534.2006

FIG. 2.

FIG. 2.

Prp43p is associated with intronic lariat RNA, spliceosomal snRNAs, several preribosomal species, and both H/ACA- and C/D-box snoRNAs. (A) Prp43p is associated with ACT1 intronic RNA in extracts lacking Dbr1p. A 1/10 equivalent of total RNA (T) and affinity-purified RNA (P) prepared from extracts of wild-type (WT), PRP43-TAP, and PRP43-TAP/Δdbr1 cells was separated by urea-PAGE, transferred to nylon membranes, and Northern blotted with an ACT1 intron oligonucleotide. (B) U2, U5, and U6 spliceosomal snRNAs are associated with Prp43p. RNA from extracts equivalent to 1/10 of the applied affinity-purified material (T) and immunopurified (P) RNAs from wild-type (WT) and Prp43p-TAP extracts were analyzed by primer extension reactions using oligonucleotides specific to the U1, U2, U4, U5, and U6 snRNAs. Primer extension products were resolved by urea-PAGE. (C) Prp43p-TAP immunopurifies several pre-rRNA processing intermediates. A 1/10 equivalent of total RNA (T) and immunoglobulin G-purified RNAs from WT and Prp43p-TAP extracts was analyzed by primer extension or RNase protection analysis as described in Materials and Methods. Products were resolved by urea-PAGE. (D) Mature rRNA genes are not associated with Prp43p. A 1/10 equivalent of total RNA from extracts (T) and immunopurified (P) RNAs from wild-type (WT) and Prp43p-TAP extracts was analyzed by primer extension analysis for the 25S, 18S, and 5.8S rRNAs. Primer extension products were resolved by urea-PAGE. (E) Box C/D and box H/ACA snoRNAs are contained in Prp43p immunoprecipitations. Primer extension analysis of U14, U24, U3, and snR3 snoRNAs was performed from total RNA (T) or immunoprecipitated RNAs (P), and primer extension products were resolved by urea-PAGE. (F) Prp43p immunoprecipitation control. scR1 small RNA was analyzed from a 1/10 equivalent of total RNA from extracts (T) and immunopurified (P) RNAs from wild-type (WT) and Prp43p-TAP extracts by primer extension analysis. Primer extension products were resolved by urea-PAGE. The above-described experiments were analyzed by using a phosphorimager.