RNA splicing and in vivo expression of the intron-containing td gene of bacteriophage T4

Marlene Belfort, Joan Pedersen-Lane, Karen Ehrenman, Frederick K. Chu, Gladys F. Maley, Frank Maley, David S. McPheeters, Larry Gold

Research output: Contribution to journalArticlepeer-review

Abstract

The splice junction sequence of td mRNA from T4-infected cells has been determined (5 '4. GGU-CUA4. 3') and shown to be identical to that of the RNA ligation product encoded by the cloned gene [Belfort et al. Cell 41 (1985) 375-382]. The RNA processing functions, T4 RNA ligase, T4 polynucleotide kinase, and the host prr gene product appear not to be essential for exon ligation; neither are the host endoribonucleases RNase III, RNase P and RNase E required for intron excision. While these results are consistent with the autocatalytic splicing mechanism demonstrated in vitro [Chu et al. J. Biol. Chem. 260 (1985) 10680-10688], they leave unanswered the question of which protein(s), if any, might stimulate the in vivo reaction. Analysis of the products of the cloned td gene has led to identification of two td-encoded polypeptides, namely a polypeptide corresponding to the exon-I-coding sequence (NH2-TS), and the catalytically active thymidylate synthase (TS). Kinetic and nucleotide sequence data provide evidence that NH2-TS is the product of the primary transcript and that TS is encoded by spliced mRNA. These results suggest that splicing may provide a switch controlling the relative expression of NH2-TS and TS, two proteins with markedly different temporal appearances despite their identical transcriptional and translational start sites.

Original languageEnglish (US)
Pages (from-to)93-102
Number of pages10
JournalGene
Volume41
Issue number1
DOIs
StatePublished - 1986

Keywords

  • RNA processing
  • Splice junction sequence
  • regulatory role for splicing
  • thymidylate synthase
  • two td-encoded proteins

ASJC Scopus subject areas

  • Genetics

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