The FIP1 gene encodes a component of a yeast pre-mRNA polyadenylation factor that directly interacts with poly(A) polymerase.
We have identified an essential gene, called FIP1, encoding a 327 amino acid protein interacting with yeast poly(A) polymerase (PAP1) in the two-hybrid assay. Recombinant FIP1 protein forms a 1:1 complex with PAP1 in vitro. At 37 degrees C, a thermosensitive allele of FIP1 shows a shortening of poly(A) tails ... and a decrease in the steady-state level of actin transcripts. When assayed for 3'-end processing in vitro, fip1 mutant extracts exhibit normal cleavage activity, but fail to polyadenylate the upstream cleavage product. Polyadenylation activity is restored by adding polyadenylation factor I (PF I). Antibodies directed against FIP1 specifically recognize a polypeptide in these fractions. Coimmunoprecipitation experiments reveal that RNA14, a subunit of cleavage factor I (CF I), directly interacts with FIP1, but not with PAP1. We propose a model in which PF I tethers PAP1 to CF I, thereby conferring specificity to poly(A) polymerase for pre-mRNA substrates.
Mesh Terms:
Actins, Amino Acid Sequence, Base Sequence, Cloning, Molecular, Genes, Fungal, Models, Genetic, Molecular Sequence Data, Mutagenesis, Poly A, Polynucleotide Adenylyltransferase, Protein Binding, Protein Denaturation, RNA Precursors, RNA Processing, Post-Transcriptional, RNA, Messenger, RNA-Binding Proteins, Recombinant Fusion Proteins, Regulatory Sequences, Nucleic Acid, Saccharomyces cerevisiae, Sequence Analysis, DNA, mRNA Cleavage and Polyadenylation Factors
Actins, Amino Acid Sequence, Base Sequence, Cloning, Molecular, Genes, Fungal, Models, Genetic, Molecular Sequence Data, Mutagenesis, Poly A, Polynucleotide Adenylyltransferase, Protein Binding, Protein Denaturation, RNA Precursors, RNA Processing, Post-Transcriptional, RNA, Messenger, RNA-Binding Proteins, Recombinant Fusion Proteins, Regulatory Sequences, Nucleic Acid, Saccharomyces cerevisiae, Sequence Analysis, DNA, mRNA Cleavage and Polyadenylation Factors
Cell
Date: May. 05, 1995
PubMed ID: 7736590
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