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Updated: Jul 20, 2026

Toeprinting Analysis of Translation Initiation Complex Formation on Mammalian mRNAs
Published on: May 10, 2018
The precursor of mouse beta-globin messenger RNA contains two intervening RNA sequences
Abstract:
We have investigated the locations of the poly(A), the mRNA-specific sequences and the RNA sequences that are eventually cleaved from the 1860 nucleotide precursor of mouse beta-globin mRNA. Biochemical and electron microscopic data demonstrate that there are two intervening RNA moieties in the precursor which separate the beta-globin mRNA sequences into three portions containing 480, 205 and 155 nucleotides. One of the two intervening RNA moieties contains 780 nucleotides. The size of the smaller intervening RNA has not been determined precisely, but it is 125 nucleotides or less. The largest mRNA-specific fragment is derived from the 3' terminus of the precursor, and contains the 3' terminal poly (A) and 330 mRNA-specific transcribed nucleotides. At least one, and probably both, intervening RNAs occur within the coding portion of the mRNA sequences. The larger of the intervening RNAs is located next to the 480 nucleotide mRNA-specific fragment, and the smaller intervening RNA is located between the 205 and 155 nucleotide mRNA-specific fragments. These experiments are consistent with the notion that the intervening sequences in the DNA of mouse beta-globin genes are transcribed into the mRNA precursor and are excised from the RNA by post-transcriptional events.
Insights
Researchers mapped RNA sequences in mouse beta-globin mRNA precursors. They identified intervening RNA moieties, suggesting post-transcriptional processing is key for mature mRNA formation.
Area of Science:
- Molecular Biology
- RNA Processing
- Gene Expression
Background:
- Mouse beta-globin mRNA precursor is 1860 nucleotides long.
- Understanding the structure of precursor mRNA is crucial for deciphering gene expression regulation.
Purpose of the Study:
- To investigate the locations of poly(A) sequences, mRNA-specific sequences, and cleaved RNA fragments within the mouse beta-globin mRNA precursor.
- To determine the size and location of intervening RNA moieties.
Main Methods:
- Biochemical analysis
- Electron microscopy
Main Results:
- Two intervening RNA moieties were identified in the precursor.
- These moieties separate beta-globin mRNA sequences into 480, 205, and 155 nucleotide portions.
- The larger intervening RNA is 780 nucleotides; the smaller is ≤125 nucleotides.
- The 3' end contains poly(A) and 330 transcribed nucleotides.
- Intervening RNAs are located within the coding portion of mRNA sequences.
Conclusions:
- Experiments support the transcription of intervening DNA sequences into mRNA precursors.
- RNA excision occurs via post-transcriptional events.
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