Purification of the Saccharomyces cerevisiae cleavage/polyadenylation factor I. Separation into two components that are required for both cleavage and polyadenylation of mRNA 3' ends.
The cleavage/polyadenylation factor I (CF I) is one of four factors required for mRNA 3' end formation in the yeast Saccharomyces cerevisiae. Here we describe the purification of CF I and its separation into two components, CF IA and CF IB. Both components are needed to reconstitute CF I activity in cleavage and poly(A) addition. CF IA consists of a complex of four polypeptides of 76, 70, 50, and 38 kDa, and CF IB is a single 73-kDa polypeptide. The 76- and 38-kDa subunits of CF IA correspond to the previously identified RNA14 and RNA15 proteins. The RNA14 protein, but not the 70- or 50-kDa proteins, coimmunoprecipitates with the RNA15 protein, indicating that RNA14 and RNA15 proteins exist in a tight complex. RNA15 is the only subunit of CF I that can be cross-linked to pre-mRNA.
Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111-1800, USA.
M M
MM
Kessler
J
J
Zhao
C L
CL
Moore
D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D011061:Poly A / Q000378:metabolism; D012331:RNA, Fungal / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D039221:mRNA Cleavage and Polyadenylation Factors
D005656:Fungal Proteins; D009687:Nuclear Proteins; D012331:RNA, Fungal; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D039221:mRNA Cleavage and Polyadenylation Factors; C512638:RNA14 protein, S cerevisiae; C512635:RNA15 protein, S cerevisiae; D011061:Poly A