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. 2002 Sep;22(18):6648–6660. doi: 10.1128/MCB.22.18.6648-6660.2002

FIG. 3.

FIG. 3.

A sequence element downstream of the stem-loop is required for Drosophila histone pre-mRNA processing. (A) Sequence of the dH3∗ pre-mRNA starting first nucleotide after the stem-loop. The six mutations M1 to M6 are shown with the nucleotide substitutions indicated above and below the wild-type sequence. (B) In vitro processing of the wild-type (WT) and mutant dH3∗ pre-mRNAs (indicated above each lane) in the nuclear extract from S-2 cells. The processing of M1 and M2 and of M3 to M6 mutant pre-mRNAs was analyzed on two gels (lanes 1 to 3 and 4 to 7), accounting for the differences in separation of unprocessed and processed RNAs across the panel. (C) dNE from S-2 cells was incubated with a control monoclonal antibody (mock, lane 2) or anti-Sm antibody (lane 3), and the antibody complexes were removed from the extract with protein G beads. The processing activity of the depleted nuclear extract was assayed with dH3∗ pre-mRNA. Lane 1 represents untreated nuclear extract.