TY - JOUR
T1 - The 30-kd subunit of mammalian cleavage and polyadenylation specificity factor and its yeast homolog are rna-binding zinc finger proteins
AU - Barabino, Silvia M.L.
AU - Hübner, Wolfgang
AU - Jenny, Andreas
AU - Minvielle-Sebastia, Lionel
AU - Keller, Walter
PY - 1997/7/1
Y1 - 1997/7/1
N2 - Cleavage and polyadenylation specificity factor (CPSF), a key component of the mammalian RNA 3-end processing machinery, consists of four subunits of 160, 100, 73, and 30 kD. Here we report the isolation and characterization of a cDNA encoding the 30-kD polypeptide. Antibodies raised against this protein inhibit cleavage and polyadenylation and coimmunoprecipitate the other CPSF subunits. The protein sequence contains five C3H-zinc-finger repeats and a putative RNA-binding zinc knuckle motif at the carboxyl terminus. Consistent with this observation, the in vitro translated 30-kD protein binds RNA polymers with a distinct preference for poly(U). In addition, an essential S. cerevisiae gene, YTH1, was cloned which is 40% identical to CPSF 30K at the protein level. Extracts prepared from a conditional yth1 mutant have normal cleavage activity, but fail to polyadenylate the upstream cleavage product. Efficient polyadenylation activity can be restored by the addition of purified polyadenylation factor I (PF I). We demonstrate that Yth1p is a component of PF I that interacts in vivo and in vitro with Fip1p, a known PF I subunit.
AB - Cleavage and polyadenylation specificity factor (CPSF), a key component of the mammalian RNA 3-end processing machinery, consists of four subunits of 160, 100, 73, and 30 kD. Here we report the isolation and characterization of a cDNA encoding the 30-kD polypeptide. Antibodies raised against this protein inhibit cleavage and polyadenylation and coimmunoprecipitate the other CPSF subunits. The protein sequence contains five C3H-zinc-finger repeats and a putative RNA-binding zinc knuckle motif at the carboxyl terminus. Consistent with this observation, the in vitro translated 30-kD protein binds RNA polymers with a distinct preference for poly(U). In addition, an essential S. cerevisiae gene, YTH1, was cloned which is 40% identical to CPSF 30K at the protein level. Extracts prepared from a conditional yth1 mutant have normal cleavage activity, but fail to polyadenylate the upstream cleavage product. Efficient polyadenylation activity can be restored by the addition of purified polyadenylation factor I (PF I). We demonstrate that Yth1p is a component of PF I that interacts in vivo and in vitro with Fip1p, a known PF I subunit.
KW - Cleavage and polyadenylation specificity factor
KW - Polyadenylation factor I
KW - Pre-mRNA processing
KW - Yeast
KW - Zinc finger
KW - Zinc knuckle
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U2 - 10.1101/gad.11.13.1703
DO - 10.1101/gad.11.13.1703
M3 - Article
C2 - 9224719
AN - SCOPUS:1842329727
SN - 0890-9369
VL - 11
SP - 1703
EP - 1716
JO - Genes and Development
JF - Genes and Development
IS - 13
ER -