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Nucleic Acids Research, 2000, Vol. 28, No. 2 520-526
© 2000 Oxford University Press

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

Lee S. Hatton, Jyrki J. Eloranta, Luisa M. Figueiredo, Yoshio Takagaki1, James L. Manley1 and Kevin O’Hare*

Department of Biochemistry, Imperial College of Science, Technology and Medicine, London SW7 2AZ, UK and 1Department of Biological Sciences, Columbia University, 1117 Fairchild Center, New York, NY 10027, USA

During mRNA 3' end formation, cleavage stimulation factor (CstF) binds to a GU-rich sequence downstream from the polyadenylation site and helps to stabilise 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 binding 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 Drosophila homologue for CstF-64 has now been isolated, both through homology with the human protein and through protein–protein interaction in yeast with the suppressor of forked gene product. Alignment of CstF-64 homologues shows that the proteins have a conserved N-terminal 200 amino acids, the first half of which is the RNA binding domain with the second half likely to contain the CstF-77 interaction domain; a central region variable in length and rich in glycine, proline and glutamine residues and containing an unusual degenerate 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 throughout development and probably corresponds to l(3)91Cd.

* To whom correspondence should be addressed. Tel: +44 207 594 5292; Fax: +44 207 594 5207; Email: k.ohare@ic.ac.uk Present addresses: Jyrki J. Eloranta, Imperial Cancer Research Fund Molecular Oncology Unit, MRC Cyclotron Building, Hammersmith Hospital, London W12 0NN, UK Luisa M. Figueiredo, Unité de Biologie des Intéractions Hôte-Parasite, CNRS URA 1960, Institut Pasteur, 75724 Paris, France Yoshio Takagaki, Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville, VA 22908, USA


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