The Drosophila homologue of the 64 kDa subunit of cleavage stimulation factor interacts with the 77 kDa subunit encoded by the suppressor of forked gene

During mRNA 3' end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the polyadenylation site and helps to sta bilise the binding of cleavage-polyadenylation specificity factor (CPSF) to the upstream polyadenylation sequence (AAUAAA), The 64 kDa subunit of CstF (CstF-64) contains an RNA binding domain and is responsible for the RNA bi nding activity of CstF, It interacts with CstF-77, which in turn interacts with CPSF, The Drosophila suppressor of forked gene encodes a homologue of CstF-77, and mutations in it affect mRNA 3' end formation in vivo. A Drosop hila homologue for CstF-64 has now been isolated, both through homology wit h the human protein and through protein-protein interaction in yeast with t he suppressor of forked gene product, Alignment of CstF-64 homologues shows that the proteins have a conserved N-terminal 200 amino acids, the first h alf of which is the RNA binding domain with the second half likely to conta in the CstF-77 interaction domain; a central region variable in length and rich in glycine, proline and glutamine residues and containing an unusual d egenerate repeat motif; and then a conserved C-terminal 50 amino acids, In Drosophila, the CstF-64 gene has a single 63 bp intron, is transcribed thro ughout development and probably corresponds to I(3)91Cd.