TRF4 is involved in polyadenylation of snRNAs in Drosophila melanogaster

Mol Cell Biol. 2008 Nov;28(21):6620-31. doi: 10.1128/MCB.00448-08. Epub 2008 Sep 2.

Abstract

The Saccharomyces cerevisiae poly(A) polymerases Trf4 and Trf5 are involved in an RNA quality control mechanism, where polyadenylated RNAs are degraded by the nuclear exosome. Although Trf4/5 homologue genes are distributed throughout multicellular organisms, their biological roles remain to be elucidated. We isolated here the two homologues of Trf4/5 in Drosophila melanogaster, named DmTRF4-1 and DmTRF4-2, and investigated their biological function. DmTRF4-1 displayed poly(A) polymerase activity in vitro, whereas DmTRF4-2 did not. Gene knockdown of DmTRF4-1 by RNA interference is lethal in flies, as is the case for the trf4 trf5 double mutants. In contrast, disruption of DmTRF4-2 results in viable flies. Cellular localization analysis suggested that DmTRF4-1 localizes in the nucleolus. Abnormal polyadenylation of snRNAs was observed in transgenic flies overexpressing DmTRF4-1 and was slightly increased by the suppression of DmRrp6, the 3'-5' exonuclease of the nuclear exosome. These results suggest that DmTRF4-1 and DmRrp6 are involved in the polyadenylation-mediated degradation of snRNAs in vivo.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Nucleolus / enzymology
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / enzymology*
  • Drosophila melanogaster / genetics
  • Genes, Developmental
  • Genome, Insect / genetics
  • Molecular Sequence Data
  • Morphogenesis
  • Mutation / genetics
  • Polyadenylation*
  • Polynucleotide Adenylyltransferase / chemistry
  • Polynucleotide Adenylyltransferase / metabolism*
  • Protein Transport
  • RNA, Small Nuclear / metabolism*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism
  • Sequence Homology, Nucleic Acid
  • Subcellular Fractions / enzymology

Substances

  • Drosophila Proteins
  • RNA, Small Nuclear
  • Saccharomyces cerevisiae Proteins
  • Polynucleotide Adenylyltransferase
  • DNA-Directed DNA Polymerase
  • PAP2 protein, S cerevisiae