Exon circularization in mammalian nuclear extracts

Zvi Pasman, Michael D. Been, Mariano Garcia-Blanco

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Correct ligation of exons in pre-mRNA splicing requires splice site juxtaposition (splice site pairing), usually involving a 5' splice site and a downstream 3' splice site. Splicing of a 5' splice site to an upstream 3' splice site, however, is predicted to result in a circular RNA. This mode of splice site pairing across the exon has been hypothesized to account for rare RNAs containing scrambled exons (Nigro JM et al., 1991, Cell 64:607-613; Cocquerelle C et al., 1992, EMBO J11:1095-1098). Additionally, this mode of splice site pairing has been postulated to explain the formation of SRY circular transcripts in mouse testis (Capel Bet al., 1993, Cell 73:1019- 1030). Here we show that splice site pairing across the exon can result in exon circularization in vitro. These results indicate that spliceosome- mediated exon circularization indeed can account for the formation of scrambled exons and circular RNAs. Exon circularization efficiency decreased dramatically as the length of the exon was increased from 95 nt to 274 nt. Circularization of this longer exon was restored, however, when intronic complementary sequences were included in the RNA substrate. These complementary sequences could form a stem that served to bring the splice sites into proximity and thereby promote splice site pairing. Therefore, the splicing of this structured RNA recapitulated SRY-like exon circularization in vitro.

Original languageEnglish (US)
Pages (from-to)603-610
Number of pages8
JournalRNA
Volume2
Issue number6
StatePublished - 1996
Externally publishedYes

Fingerprint

Exons
RNA Splice Sites
RNA
Spliceosomes
RNA Splicing
RNA Precursors
Ligation
Testis

Keywords

  • circular exons
  • circularization
  • pre-mRNA splicing

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Pasman, Z., Been, M. D., & Garcia-Blanco, M. (1996). Exon circularization in mammalian nuclear extracts. RNA, 2(6), 603-610.

Exon circularization in mammalian nuclear extracts. / Pasman, Zvi; Been, Michael D.; Garcia-Blanco, Mariano.

In: RNA, Vol. 2, No. 6, 1996, p. 603-610.

Research output: Contribution to journalArticle

Pasman, Z, Been, MD & Garcia-Blanco, M 1996, 'Exon circularization in mammalian nuclear extracts', RNA, vol. 2, no. 6, pp. 603-610.
Pasman, Zvi ; Been, Michael D. ; Garcia-Blanco, Mariano. / Exon circularization in mammalian nuclear extracts. In: RNA. 1996 ; Vol. 2, No. 6. pp. 603-610.
@article{0d1832290bf24391902d751a000322a5,
title = "Exon circularization in mammalian nuclear extracts",
abstract = "Correct ligation of exons in pre-mRNA splicing requires splice site juxtaposition (splice site pairing), usually involving a 5' splice site and a downstream 3' splice site. Splicing of a 5' splice site to an upstream 3' splice site, however, is predicted to result in a circular RNA. This mode of splice site pairing across the exon has been hypothesized to account for rare RNAs containing scrambled exons (Nigro JM et al., 1991, Cell 64:607-613; Cocquerelle C et al., 1992, EMBO J11:1095-1098). Additionally, this mode of splice site pairing has been postulated to explain the formation of SRY circular transcripts in mouse testis (Capel Bet al., 1993, Cell 73:1019- 1030). Here we show that splice site pairing across the exon can result in exon circularization in vitro. These results indicate that spliceosome- mediated exon circularization indeed can account for the formation of scrambled exons and circular RNAs. Exon circularization efficiency decreased dramatically as the length of the exon was increased from 95 nt to 274 nt. Circularization of this longer exon was restored, however, when intronic complementary sequences were included in the RNA substrate. These complementary sequences could form a stem that served to bring the splice sites into proximity and thereby promote splice site pairing. Therefore, the splicing of this structured RNA recapitulated SRY-like exon circularization in vitro.",
keywords = "circular exons, circularization, pre-mRNA splicing",
author = "Zvi Pasman and Been, {Michael D.} and Mariano Garcia-Blanco",
year = "1996",
language = "English (US)",
volume = "2",
pages = "603--610",
journal = "RNA",
issn = "1355-8382",
publisher = "Cold Spring Harbor Laboratory Press",
number = "6",

}

TY - JOUR

T1 - Exon circularization in mammalian nuclear extracts

AU - Pasman, Zvi

AU - Been, Michael D.

AU - Garcia-Blanco, Mariano

PY - 1996

Y1 - 1996

N2 - Correct ligation of exons in pre-mRNA splicing requires splice site juxtaposition (splice site pairing), usually involving a 5' splice site and a downstream 3' splice site. Splicing of a 5' splice site to an upstream 3' splice site, however, is predicted to result in a circular RNA. This mode of splice site pairing across the exon has been hypothesized to account for rare RNAs containing scrambled exons (Nigro JM et al., 1991, Cell 64:607-613; Cocquerelle C et al., 1992, EMBO J11:1095-1098). Additionally, this mode of splice site pairing has been postulated to explain the formation of SRY circular transcripts in mouse testis (Capel Bet al., 1993, Cell 73:1019- 1030). Here we show that splice site pairing across the exon can result in exon circularization in vitro. These results indicate that spliceosome- mediated exon circularization indeed can account for the formation of scrambled exons and circular RNAs. Exon circularization efficiency decreased dramatically as the length of the exon was increased from 95 nt to 274 nt. Circularization of this longer exon was restored, however, when intronic complementary sequences were included in the RNA substrate. These complementary sequences could form a stem that served to bring the splice sites into proximity and thereby promote splice site pairing. Therefore, the splicing of this structured RNA recapitulated SRY-like exon circularization in vitro.

AB - Correct ligation of exons in pre-mRNA splicing requires splice site juxtaposition (splice site pairing), usually involving a 5' splice site and a downstream 3' splice site. Splicing of a 5' splice site to an upstream 3' splice site, however, is predicted to result in a circular RNA. This mode of splice site pairing across the exon has been hypothesized to account for rare RNAs containing scrambled exons (Nigro JM et al., 1991, Cell 64:607-613; Cocquerelle C et al., 1992, EMBO J11:1095-1098). Additionally, this mode of splice site pairing has been postulated to explain the formation of SRY circular transcripts in mouse testis (Capel Bet al., 1993, Cell 73:1019- 1030). Here we show that splice site pairing across the exon can result in exon circularization in vitro. These results indicate that spliceosome- mediated exon circularization indeed can account for the formation of scrambled exons and circular RNAs. Exon circularization efficiency decreased dramatically as the length of the exon was increased from 95 nt to 274 nt. Circularization of this longer exon was restored, however, when intronic complementary sequences were included in the RNA substrate. These complementary sequences could form a stem that served to bring the splice sites into proximity and thereby promote splice site pairing. Therefore, the splicing of this structured RNA recapitulated SRY-like exon circularization in vitro.

KW - circular exons

KW - circularization

KW - pre-mRNA splicing

UR - http://www.scopus.com/inward/record.url?scp=0029944946&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029944946&partnerID=8YFLogxK

M3 - Article

C2 - 8718689

AN - SCOPUS:0029944946

VL - 2

SP - 603

EP - 610

JO - RNA

JF - RNA

SN - 1355-8382

IS - 6

ER -