Journalartikel

Jahr der Veröffentlichung: 1990

Seiten: 201-224

Zeitschrift: Genetic Engineering: Principles and Methods

Bandnummer: 12

ISSN: 0196-3716

DOI Link: https://doi.org/10.1007/978-1-4613-0641-2_11

Verlag: Springer

Abstract: 

In eukaryotic cells most primary transcripts of protein-coding genes are
processed through a variety of post-transcriptional modification
reactions to yield mature mRNA. One of these processing reactions termed
pre-mRNA splicing involves the accurate excision of intron RNA
sequences and the coordinate ligation of exon RNA sequences. In
addition, there are post-transcriptional modifications at the 5’ end
(capping) and at the 3’ end (polyadenylation). A systematic biochemical
study of premRNA splicing began with the development of efficient in vitro
splicing systems [reviewed in (1)] that accurately splice synthetic
pre-mRNAs in nuclear extracts prepared from cultured mammalian cells. In
most cases, relatively short model substrates consisting of two exons
and one intron were used for these studies (see Figure 1 for a schematic
representation). The biochemical analysis of this highly complex RNA
processing reaction has since made rapid progress. Initial studies
revealed a two-step mechanism involving the three conserved splicing
signals (Figure 1). First, cleavage at the 5’ splice site occurs
concomitantly with lariat formation at the intron branch point. Second,
the RNA intermediates, exon 1 and intron lariat-exon 2, are cleaved at
the 3’ splice site, and the two exon RNA sequences are ligated together.
The intron RNA sequences are thereby released in the form of a lariat.
Soon after the establishment of in vitro
splicing systems it became clear that this complex series of reactions
occurs in large ribonucleoprotein (RNP) structures of the pre-mRNA
called splicing complexes or spliceo­somes (2–7). The ordered assembly
of splicing complexes is a prerequisite of the pre-mRNA splicing
reaction and requires the coordinate action of small nuclear
ribonucleoproteins (snRNPs) and of additional proteins, so-called
non-snRNP splicing factors.

Harvard-Zitierstil: Bindereif, A. and Green, M. (1990) Identification and functional analysis of mammalian splicing factors., Genetic Engineering: Principles and Methods, 12, pp. 201-224. https://doi.org/10.1007/978-1-4613-0641-2_11

APA-Zitierstil: Bindereif, A., & Green, M. (1990). Identification and functional analysis of mammalian splicing factors.. Genetic Engineering: Principles and Methods. 12, 201-224. https://doi.org/10.1007/978-1-4613-0641-2_11