A single-base change at a splice site in a β0-thalassemic gene causes abnormal RNA splicing

Treisman, Richard; Proudfoot, Nick J.; Shander, Monica H. M.; Maniatis, Tom

doi:10.1016/0092-8674(82)90452-4

Cited by 357 publications

(159 citation statements)

References 35 publications

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“…PCR conditions for the direct amplification of genomic DNA or RNA without reverse transcription were exactly the same as for RT-PCR. Reverse transcriptase mapping for the determination of the transcription initiation site was carried out basically as described by Treisman et al (1982).…”

Section: Analysis Of Rna Transcriptsmentioning

confidence: 99%

A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization

Uemori¹,

Sato²,

Kato³

et al. 1997

Genes to Cells

117

140

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Background: In many respects Archaea are much more like eukaryotes than prokaryotes with respect to the conservation of many of the components involved in transcription, translation and DNA replication. So far, only a few DNA polymerases with structures similar to those of eukaryotic DNA polymerase a have been found in Archaea. The identification and characterization of all the DNA polymerases of one archaeon would add considerably to our knowledge of the basic mechanisms of DNA replication in these organisms.

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Section: Analysis Of Rna Transcriptsmentioning

confidence: 99%

A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization

Uemori¹,

Sato²,

Kato³

et al. 1997

Genes to Cells

117

140

View full text Add to dashboard Cite

show abstract

“…On the other hand, mutants have been described where alternative splicing occurs, but apparently is not very efficient. In such cases unspliced precursor accumulates (4,44). Results with these mutants have suggested that there is a strong propensity for RNA to be spliced, as it occurs even on elimination of a wild type splice site.…”

Section: Discussiqnmentioning

confidence: 99%

“…One fate is that efficient splicing can occur, using alternative splice sites. These new splice sites can be wild type sites (in the case of overlapping genes) or previously unused, cryptic sites (1,2,4,7,43). On the other hand, mutants have been described where alternative splicing occurs, but apparently is not very efficient.…”

Section: Discussiqnmentioning

confidence: 99%

“…Analyses of intron sequence requirements for in vivo mRNA splicing in higher eukaryotes originally indicated the importance of the 5' splice site, including the highly conserved GU dinucleotide at the 5' exon-intron boundary, and the 3' splice site, which consists of a similarly highly conserved dinucleotide, AG, at the 3' boundary, but which additionally includes an upstream stretch of pyrimidines (1)(2)(3)(4)(5)(6)(7). More recently, a third site which shows some sequence requirements for splicing in vivo has been identified -the branch point, which lies about 30 nucleotides upstream of the 3' splice site (8).…”

Section: Introductionmentioning

confidence: 99%

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The length but not the sequence of the polyoma virus late leader exon is important for both late RNA splicing and stability

Adami¹,

Carmichael

1987

Nucl Acids Res

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ABSTRACT'Polyoma virus late RNA processing provides a convenient model system in which to study the mechanics of splicing in vivo. In order to understand further the role of the untranslated "late leader" unit in late RNA processing we have constructed a group of polyoma viruses with deletions and substitutions in the leader exon. This has allowed us to determine that there is a minimum exon size required for both pre-mRNA splicing and stability in this system. We show here that the non-viability of a mutant (ALM) with a 9 base late leader unit is due to a general defect in late RNA splicing. In addition, ALM-infected cells show at least 40-fold depression in the accumulation of late nuclear RNA (spliced or unspliced). The ALM late promoter, however, functions nearly normally. Substituted leader variants with 51-to 96-base long exons of unrelated sequence are viable (G. Adami and G. Carmichael, J. Virol. 58, 417-425, 1986). We show here that late RNA from one of these substituted leader mutants (containing a 51-base leader exon) is spliced at wild type levels, with virtually no defect in accumulation. Thus, in the polyoma system, splice sites separated by only 9 bases can inhibit each others usage, presumably by steric interference. We suggest that this type of inhibition leads to extreme RNA instability. INTRODUCTION Much work in recent years has been directed towards further understanding the mechanism of splicing of mRNA precursors. Analyses of intron sequence requirements for in vivo mRNA splicing in higher eukaryotes originally indicated the importance of the 5' splice site, including the highly conserved GU dinucleotide at the 5' exon-intron boundary, and the 3' splice site, which consists of a similarly highly conserved dinucleotide, AG, at the 3' boundary, but which additionally includes an upstream stretch of pyrimidines (1-7). More recently, a third site which shows some sequence requirements for splicing in vivo has been identified -the branch point,

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“…Polar effects seem to be a general feature of the many splicing mutations discovered: they have been reported for nuclear genes [50,51] and for the yeast mitochondrial cob-box gene (vide supra). The oxi3 case would thus seem to be an exception.…”

Section: Results)mentioning

confidence: 99%

Molecular basis of the ‘box effect’

Labouesse

Netter

Schroeder

1984

European Journal of Biochemistry

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In the mitochondrial DNA of Saccharomyces cerevisiue, the genes cob-box and oxi3, coding for apocytochrome b and cytochrome oxidase subunit I respectively, are split. Several mutations located in the introns of the cob-box gene prevent the synthesis of cytochrome b and cytochrome oxidase subunit I (this is known as the 'box effect').-We have elucidated the molecular basis of this phenomenon : these mutants are unable to excise the fourth intron of oxi3 from the cytochrome oxidase subunit 1 pre-mRNA; the absence of a functional b14 mRNA maturase, a trans-acting factor encoded by the fourth intron of the cob-box gene explains this phenomenon. This maturase was already known to control the excision of the b14 intron; consequently we have demonstrated that it is necessary for the processing of two introns located in two different genes.Mutations altering this maturase can be corrected, but only partially, by extragenic suppressors located in the mitochondrial (mim2) or in the nuclear (NAM2) genome. The gene product of these two suppressors should, therefore, control (directly or indirectly) the excision of the two introns as the b14 mRNA maturase normally does.In Saccharomyces cerevisiae mitochondria the genes cobbox and oxi3, coding respectively for apocytochrome b [ 1 -61 and cytochrome oxidase subunit I [4,, are split genes. Several hundred mutations mapping within exons as well as within the introns of these genes have been isolated (for a recent review see [lo]). Complementation analyses [I I], hybridization studies of mitochondrial transcripts and processing defects [ 12 -161, and DNA sequencing of mutations [6,17 -211 have revealed two classes of intron mutations. Mutations belonging to the first class exhibit cis-dominant complementation behavior. They always affect very short critical sequences located within introns (these critical sequences are homologous between different introns) and they block RNA splicing very probably by altering the substrate of the splicing reaction (i.e. RNA specific sequence and/or secondary structures required for processing, cf. 22 -24). Mutations belonging to the second class exhibit trans-recessive complementation behavior and are dispersed along the open reading frames present in some introns (the 2nd, 3rd and 4th of cob-box gene, the 1st of oxi3 gene). They block RNA processing by modifying the activity of a protein component of the splicing machinery, thought to be required for the excision of the intron which encodes it. These proteins have been called the mRNA maturases [6,17,19,20,25] (Lazowska et al. unpublished results).The expression of the genes cob-box and oxi3 is linked by a particular interaction : several mutations located in the cobbox gene prevent cytochrome oxidase subunit I synthesis. _ _ _ _ b14 maturase (coded by the 4th intron of the cob-box gene) is necessary for oxi3 gene expression and thus responsible for the box effect; (a) only trans-recessive mutations in the 4th intron of the cob-box gene (b14 intron) display this effect while cisdominant ones do...

show abstract

A single-base change at a splice site in a β0-thalassemic gene causes abnormal RNA splicing

Cited by 357 publications

References 35 publications

A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization

A novel DNA polymerase in the hyperthermophilic archaeon, Pyrococcus furiosus: gene cloning, expression, and characterization

The length but not the sequence of the polyoma virus late leader exon is important for both late RNA splicing and stability

Molecular basis of the ‘box effect’

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