β-globin transcripts carrying a single intron with three adjacent nucleotides of 5′ exon are efficiently spliced in vitro irrespective of intron position or surrounding exon sequences

Akira Maeda, Yasumi Ohshima

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

To examine the role of exon sequences and intron position in the splicing of an mRNA precursor, we prepared series of sense or anti-sense transcripts of human β-globin cDNA in which a cassette containing the β-globin first intron was inserted into one of seven unusual positions. The intron cassette consisted of the intron alone (MI), the intron with three adjacent base pairs of the 5′ exon (MI), or the intron with both 5′ and 3′ exon sequences. All these transcripts were examined in an in vitro splicing system with a HeLa cell nuclear extract. The sense transcripts carrying MI cassette were spliced efficiently and independently of the intron position, except when the 3′ exon was too short. The anti-sense transcripts carrying MI cassette produced significantly less spliced products than did those of the sense transcripts. This was mostly because of the instability of the anti-sense transcripts, and the actual splicing efficiency was similar to that seen in the sense transcripts. Sense or anti-sense transcripts carrying MI cassette were spliced to various extents depending on the surrounding sequences. The results indicate that only three nucleotides of the 5′ exon are required as specific exon sequences in the splicing of an mRNA precursor carrying a single intron, and that the intron position does not significantly affect the splicing efficiency in vitro.

Original languageEnglish
Pages (from-to)4671-4676
Number of pages6
JournalNucleic Acids Research
Volume18
Issue number16
DOIs
Publication statusPublished - 25-08-1990

Fingerprint

Globins
Nucleotides
Introns
Exons
Adjacent
Precursor
Messenger RNA
CDNA
RNA Precursors
Series
In Vitro Techniques
Cell
Cell Extracts
HeLa Cells
Base Pairing
Complementary DNA

All Science Journal Classification (ASJC) codes

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

Cite this

@article{b92a9895ae4746f2a03d4900aa29de05,
title = "β-globin transcripts carrying a single intron with three adjacent nucleotides of 5′ exon are efficiently spliced in vitro irrespective of intron position or surrounding exon sequences",
abstract = "To examine the role of exon sequences and intron position in the splicing of an mRNA precursor, we prepared series of sense or anti-sense transcripts of human β-globin cDNA in which a cassette containing the β-globin first intron was inserted into one of seven unusual positions. The intron cassette consisted of the intron alone (MI), the intron with three adjacent base pairs of the 5′ exon (MI), or the intron with both 5′ and 3′ exon sequences. All these transcripts were examined in an in vitro splicing system with a HeLa cell nuclear extract. The sense transcripts carrying MI cassette were spliced efficiently and independently of the intron position, except when the 3′ exon was too short. The anti-sense transcripts carrying MI cassette produced significantly less spliced products than did those of the sense transcripts. This was mostly because of the instability of the anti-sense transcripts, and the actual splicing efficiency was similar to that seen in the sense transcripts. Sense or anti-sense transcripts carrying MI cassette were spliced to various extents depending on the surrounding sequences. The results indicate that only three nucleotides of the 5′ exon are required as specific exon sequences in the splicing of an mRNA precursor carrying a single intron, and that the intron position does not significantly affect the splicing efficiency in vitro.",
author = "Akira Maeda and Yasumi Ohshima",
year = "1990",
month = "8",
day = "25",
doi = "10.1093/nar/18.16.4671",
language = "English",
volume = "18",
pages = "4671--4676",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "16",

}

TY - JOUR

T1 - β-globin transcripts carrying a single intron with three adjacent nucleotides of 5′ exon are efficiently spliced in vitro irrespective of intron position or surrounding exon sequences

AU - Maeda, Akira

AU - Ohshima, Yasumi

PY - 1990/8/25

Y1 - 1990/8/25

N2 - To examine the role of exon sequences and intron position in the splicing of an mRNA precursor, we prepared series of sense or anti-sense transcripts of human β-globin cDNA in which a cassette containing the β-globin first intron was inserted into one of seven unusual positions. The intron cassette consisted of the intron alone (MI), the intron with three adjacent base pairs of the 5′ exon (MI), or the intron with both 5′ and 3′ exon sequences. All these transcripts were examined in an in vitro splicing system with a HeLa cell nuclear extract. The sense transcripts carrying MI cassette were spliced efficiently and independently of the intron position, except when the 3′ exon was too short. The anti-sense transcripts carrying MI cassette produced significantly less spliced products than did those of the sense transcripts. This was mostly because of the instability of the anti-sense transcripts, and the actual splicing efficiency was similar to that seen in the sense transcripts. Sense or anti-sense transcripts carrying MI cassette were spliced to various extents depending on the surrounding sequences. The results indicate that only three nucleotides of the 5′ exon are required as specific exon sequences in the splicing of an mRNA precursor carrying a single intron, and that the intron position does not significantly affect the splicing efficiency in vitro.

AB - To examine the role of exon sequences and intron position in the splicing of an mRNA precursor, we prepared series of sense or anti-sense transcripts of human β-globin cDNA in which a cassette containing the β-globin first intron was inserted into one of seven unusual positions. The intron cassette consisted of the intron alone (MI), the intron with three adjacent base pairs of the 5′ exon (MI), or the intron with both 5′ and 3′ exon sequences. All these transcripts were examined in an in vitro splicing system with a HeLa cell nuclear extract. The sense transcripts carrying MI cassette were spliced efficiently and independently of the intron position, except when the 3′ exon was too short. The anti-sense transcripts carrying MI cassette produced significantly less spliced products than did those of the sense transcripts. This was mostly because of the instability of the anti-sense transcripts, and the actual splicing efficiency was similar to that seen in the sense transcripts. Sense or anti-sense transcripts carrying MI cassette were spliced to various extents depending on the surrounding sequences. The results indicate that only three nucleotides of the 5′ exon are required as specific exon sequences in the splicing of an mRNA precursor carrying a single intron, and that the intron position does not significantly affect the splicing efficiency in vitro.

UR - http://www.scopus.com/inward/record.url?scp=0024999326&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024999326&partnerID=8YFLogxK

U2 - 10.1093/nar/18.16.4671

DO - 10.1093/nar/18.16.4671

M3 - Article

C2 - 1697673

AN - SCOPUS:0024999326

VL - 18

SP - 4671

EP - 4676

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 16

ER -