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Six fractions derived from a HeLa cell nuclear extract are necessary for the generation of spliced mRNA in vitro. To establish a function for the protein factors present in these fractions, their role in the formation of splicing complexes was analyzed by electrophoresis in native polyacrylamide gels. Two of the fractions are sufficient to assemble the adenovirus major late mRNA precursor into a presplicing complex with characteristics similar to the presplicing complex assembled in nuclear extract. One fraction supplies splicing factor (SF) 1 and at least one small nuclear ribonucleoprotein particle, U2 snRNP. The other fraction contains SF3. Extensive fractionation of this protein has revealed that it is essential for presplicing complex assembly and the splicing reaction.

Presplicing complex formation requires two proteins and U2 snRNP.