A polyadenylation factor subunit is the human homologue of the Drosophila suppressor of forked protein.
Polyadenylation of messenger RNA precursors is a complex process that requires multiple protein factors (for reviews, see refs 1, 2). Cleavage stimulation factor (CstF) is one of these, functioning together with cleavage-polyadenylation specificity factor, two cleavage factors, and poly(A)+ polymerase. CstF is composed of three subunits of M(r) 77, 64 and 50K. The 64K and 50K subunits contain, respectively, an RNP-type RNA-binding domain that contacts the pre-mRNA and transducin repeats characteristic of G-protein beta-subunits. Here we report the cloning and characterization of the 77K subunit of human CstF (referred to as 77K). We show that the 77K subunit is required for formation of active CstF and bridges the 64K and 50K subunits. Sequence analyses indicate that the 77K subunit is the homologue of the protein encoded by the Drosophila melanogaster suppressor of forked (su(f)) gene. Mutations in su(f) can enhance or suppress the effects of transposable element insertions, and our data indicate that this is due to changes in polyadenylation. Both the 77K subunit and the su(f) protein share homology with Saccharomyces cerevisiae RNA14, previously shown to be involved in mRNA metabolism. Our results thus also indicate that components of the complex polyadenylation machinery are conserved from yeast to man.
Takagaki
Y
Y
Department of Biological Sciences, Columbia University, New York, New York 10027.
Manley
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D039241:Cleavage Stimulation Factor / Q000737:chemistry* / Q000502:physiology; D018076:DNA, Complementary; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007120:Immunochemistry; D007301:Insect Hormones / Q000737:chemistry*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010446:Peptide Fragments / Q000737:chemistry / Q000502:physiology; D011233:Precipitin Tests; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D039221:mRNA Cleavage and Polyadenylation Factors
C546606:CSTF3 protein, human; D039241:Cleavage Stimulation Factor; D018076:DNA, Complementary; D029721:Drosophila Proteins; D007301:Insect Hormones; D009687:Nuclear Proteins; D010446:Peptide Fragments; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D039221:mRNA Cleavage and Polyadenylation Factors; C079745:su(f) protein, Drosophila