multisubunit 3' end processing factor from yeast containing poly(A) polymerase and homologues of the subunits of mammalian cleavage and polyadenylation specificity factor
Polyadenylation is the second step in 3′ end formation of most eukaryotic mRNAs. In Saccharomyces cerevisiae , this step requires three trans ‐acting factors: poly(A) polymerase (Pap1p), cleavage factor I (CF I) and polyadenylation factor I (PF I). Here, we describe the purification and subunit comp...
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Veröffentlicht in: | The EMBO journal 1997-08, Vol.16 (15), p.4727-4737 |
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Sprache: | eng |
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Zusammenfassung: | Polyadenylation is the second step in 3′ end formation of most eukaryotic mRNAs. In
Saccharomyces cerevisiae
, this step requires three
trans
‐acting factors: poly(A) polymerase (Pap1p), cleavage factor I (CF I) and polyadenylation factor I (PF I). Here, we describe the purification and subunit composition of a multiprotein complex containing Pap1p and PF I activities. PF I–Pap1p was purified to homogeneity by complementation of extracts mutant in the Fip1p subunit of PF I. In addition to Fip1p and Pap1p, the factor comprises homologues of all four subunits of mammalian cleavage and polyadenylation specificity factor (CPSF), as well as Pta1p, which previously has been implicated in pre‐tRNA processing, and several as yet uncharacterized proteins. As expected for a PF I subunit,
pta1‐1
mutant extracts are deficient for polyadenylation
in vitro
. PF I also appears to be functionally related to CPSF, as it polyadenylates a substrate RNA more efficiently than Pap1p alone. Possibly, the observed interaction of the complex with RNA tethers Pap1p to its substrate. |
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ISSN: | 0261-4189 1460-2075 1460-2075 |
DOI: | 10.1093/emboj/16.15.4727 |