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PMID: 3317279 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

Nucleic acids research ·Vol. 15 ·No. 22 ·1987-11-25 ·Pages 9279-98

Wollenzien PL, Goswami P, Teare J, Szeberenyi J, Goldenberg CJ

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 degrees 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.

MeSH Terms
Animals Base Sequence Cell Line Cell Nucleus/metabolism Cross-Linking Reagents Furocoumarins Genes HeLa Cells/metabolism Humans Insulin/genetics Mice Molecular Sequence Data Nucleic Acid Conformation Plasmacytoma RNA Precursors/genetics RNA Splicing Transcription, Genetic Trioxsalen/analogs & derivatives
Chemicals
Cross-Linking Reagents Furocoumarins Insulin RNA Precursors aminomethyltrioxsalen Trioxsalen
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wollenzien P L
Department of Microbiology and Immunology, University of Miami School of Medicine, FL 33101.
Goswami P
Teare J
Szeberenyi J
Goldenberg C J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.22.9279
Published
1987-11-25
Pages
9279-98
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306468
Subset
IM
Grants
NIAID NIH HHS · AI-24479 · United States
NIGMS NIH HHS · GM 35410 · United States
External Links
PubMed source DOI: 10.1093/nar/15.22.9279 PMC Full Text View journal details
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