The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction

Paul L. Wollenzien, Prabhat Goswami, John Teare, J. Szeberényi, Carlos J. Goldenberg

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The secondary structure of the SP6/mouse insulin precursor RNA was determined by psoralen cross-linking experiments. A series of long-range contacts occur within the left half of the pre-mRNA that contains the intervening sequence. Multiple secondary structures for the pre-mRNA exist since some of the interactions share common sites. In splicing buffer but without the splicing extract added, many of these interactions are stable up to at least 50°C. These interactions, however, are dissociated during the in vitro splicing reaction. This dissociation requires ATP and it occurs during the first 30 min, of the splicing reaction. Pre-mRNAs containing psoralen cross-links in different locations within the RNA molecule were purified and used as substrates for in vitro splicing. Psoralen cross-links at any of the double-stranded regions resulted in complete inhibition of the splicing reaction. This indicates that destabilization of the secondary structure of the SP6/mouse insulin pre-mRNA is necessary for in vitro splicing.

Original languageEnglish
Pages (from-to)9279-9298
Number of pages20
JournalNucleic Acids Research
Volume15
Issue number22
DOIs
Publication statusPublished - Nov 25 1987

Fingerprint

Ficusin
RNA Precursors
Secondary Structure
Precursor
Messenger RNA
Insulin
RNA
Mouse
Interaction
Adenosinetriphosphate
Linking
Buffer
Introns
Substrate
Molecules
Contact
Buffers
Adenosine Triphosphate
Necessary
Series

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction. / Wollenzien, Paul L.; Goswami, Prabhat; Teare, John; Szeberényi, J.; Goldenberg, Carlos J.

In: Nucleic Acids Research, Vol. 15, No. 22, 25.11.1987, p. 9279-9298.

Research output: Contribution to journalArticle

Wollenzien, Paul L. ; Goswami, Prabhat ; Teare, John ; Szeberényi, J. ; Goldenberg, Carlos J. / The secondary structure of a messenger RNA precursor probed with psoralen is melted in an in vitro splicing reaction. In: Nucleic Acids Research. 1987 ; Vol. 15, No. 22. pp. 9279-9298.
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