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In vitro reconstitution of a 3′-end processing reaction with purified factors. Lane 1, DNA markers (in nucleotides). Lane 2, unreacted CYC1 precursor. Purified CF IA [poly(U)-Sepharose fraction 34], partially purified CF II/CF IB, and purified PF I/Pap1p were incubated with CYC1 either alone (lanes 3, 4, 6, respectively) or together (CF IA + CF II/CF IB + PF I/Pap1p, lane 7). CYC1, precursor; 5′, upstream fragment; pA60–70, pAn, polyadenylated transcripts.
