The interaction between the first and last intron nucleotides in the second step of pre-mRNA splicing is independent of other conserved intron nucleotides

Ruis, Brian L.; Kivens, Wendy J.; Siliciano, Paul G.

doi:10.1093/nar/22.24.5190

Cited by 20 publications

(25 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All other pairs appeared to be inactive. These first experiments were carried out in vivo in Saccharomyces cerevisiae (Parker and Siliciano 1993;Chanfreau et al 1994;Ruis et al 1994). Subsequent investigations in mammalian in vitro systems largely confirmed the in vivo yeast findings (Deirdre et al 1995;Tarn 1996).…”

Section: Introductionmentioning

confidence: 77%

“…The only notable exception is the low frequency occurrence of U2-dependent 5 0 splice sites with a +2 C residue. Most mutations of these positions have been shown to be highly defective for splicing in U2-dependent introns (Reed and Maniatis 1985;Aebi et al 1986;Ruis et al 1994;Gaur et al 2000). We have shown previously that mutation of the 3 0 splice site À2 A residue in a U12-dependent intron blocks splicing in vivo (Dietrich et al 1997(Dietrich et al , 2001).…”

Section: In Vivo Splicing Pattern Of Mutants Of the Terminal Intron Nmentioning

confidence: 91%

“…In the case of U2-dependent introns, previous investigations of the effects of mutations in the U of the 5 0 /GU and the A residue of the 3 0 AG/ dinucleotides showed that these residues were important for efficient splicing in vivo and in vitro (Reed and Maniatis 1985;Aebi et al 1986;Ruis et al 1994;Gaur et al 2000). Unlike the case of the terminal G residues, no combinations of 5 0 and 3 0 mutations of the U and A residues restored function (Ruis et al 1994). Nevertheless, the potential for an A-U base pair interaction obviously exists and similar interactions appear to play a role in splicing of group II autocatalytic introns (Chanfreau and Jacquier 1993).…”

Section: Introductionmentioning

confidence: 99%

“…One possible interaction could involve a Watson-Crick base pair between the conserved U at +2 and the A at À2. While an analysis of an S. cerevisiae U2-type intron failed to uncover a base-pairing interaction between the +2 and À2 positions (Ruis et al 1994), the critical involvement of many other base-pairing interactions in U12-type intron splicing motivated us to examine this issue in our mammalian U12-type in vivo splicing system.…”

Section: In Vivo Splicing Pattern Of Mutants Of the Terminal Intron Nmentioning

confidence: 99%

See 3 more Smart Citations

A mutational analysis of U12-dependent splice site dinucleotides

Dietrich¹,

Fuller²,

Padgett³

2005

RNA

View full text Add to dashboard Cite

Introns spliced by the U12-dependent minor spliceosome are divided into two classes based on their splice site dinucleotides. The /AU-AC/ class accounts for about one-third of U12-dependent introns in humans, while the /GU-AG/ class accounts for the other two-thirds. We have investigated the in vivo and in vitro splicing phenotypes of mutations in these dinucleotide sequences. A 5 0 A residue can splice to any 3 0 residue, although C is preferred. A 5 0 G residue can splice to 3 0 G or U residues with a preference for G. Little or no splicing was observed to 3 0 A or C residues. A 5 0 U or C residue is highly deleterious for U12-dependent splicing, although some combinations, notably 5 0 U to 3 0 U produced detectable spliced products. The dependence of 3 0 splice site activity on the identity of the 5 0 residue provides evidence for communication between the first and last nucleotides of the intron. Most mutants in the second position of the 5 0 splice site and the next to last position of the 3 0 splice site were defective for splicing. Double mutants of these residues showed no evidence of communication between these nucleotides. Varying the distance between the branch site and the 3 0 splice site dinucleotide in the /GU-AG/ class showed that a somewhat larger range of distances was functional than for the /AU-AC/ class. The optimum branch site to 3 0 splice site distance of 11-12 nucleotides appears to be the same for both classes.

show abstract

Section: Introductionmentioning

confidence: 77%

Section: In Vivo Splicing Pattern Of Mutants Of the Terminal Intron Nmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Section: In Vivo Splicing Pattern Of Mutants Of the Terminal Intron Nmentioning

confidence: 99%

See 2 more Smart Citations

A mutational analysis of U12-dependent splice site dinucleotides

Dietrich¹,

Fuller²,

Padgett³

2005

RNA

View full text Add to dashboard Cite

show abstract

“…However, there are no previous data implicating 5ЈSS position 2 in an interaction with any residues in the 3ЈSS YAG. In fact, evidence for an interaction with the 3ЈSS penultimate residue has been sought, but not found (Ruis et al 1994), conceivably due to additional constraints imposed by Prp8.…”

Section: Allele Specificity Of 5јss and 3јss Suppressionmentioning

confidence: 99%

Allele-specific genetic interactions between Prp8 and RNA active site residues suggest a function for Prp8 at the catalytic core of the spliceosome

Collins¹,

Guthrie²

1999

Genes & Development

111

122

View full text Add to dashboard Cite

The highly conserved spliceosomal protein Prp8 is known to cross-link the critical sequences at both the 5 (GU) and 3 (YAG) ends of the intron. We have identified prp8 mutants with the remarkable property of suppressing exon ligation defects due to mutations in position 2 of the 5 GU, and all positions of the 3 YAG. The prp8 mutants also suppress mutations in position A51 of the critical ACAGAG motif in U6 snRNA, which has been observed previously to cross-link position 2 of the 5 GU. Other mutations in the 5 splice site, branchpoint, and neighboring residues of the U6 ACAGAG motif are not suppressed. Notably, the suppressed residues are specifically conserved from yeast to man, and from U2-to U12-dependent spliceosomes. We propose that Prp8 participates in a previously unrecognized tertiary interaction between U6 snRNA and both the 5 and 3 ends of the intron. This model suggests a mechanism for positioning the 3 splice site for catalysis, and assigns a fundamental role for Prp8 in pre-mRNA splicing. The removal of introns from eukaryotic genes to generate mRNA is catalyzed by the spliceosome, a complex RNA-protein machine composed of small nuclear RNAs (snRNAs), and at least 60 proteins. The pre-mRNA splicing reaction consists of two sequential transesterification steps: The first step results in cleavage of the 5Ј splice site (5ЈSS) and formation of a branched (lariat) intermediate; the second step results in ligation of the 5Ј and 3Ј exons. The existence of self-splicing group-II introns, which utilize a similar two-step transesterification mechanism, has led to the hypothesis that premRNA splicing is catalyzed by RNA (Sharp 1985;Cech 1986). Indeed, mutational and cross-linking analyses have revealed a network of RNA-RNA interactions, which have been proposed to form the catalytic core of the spliceosome (Newman 1994;Nilsen 1994).Whether any of the spliceosomal proteins make direct structural or chemical contributions to the active site remains unknown. Numerous cross-linking studies have placed one spliceosomal protein, Prp8, at or near the catalytic core. As summarized in Figure 1, Prp8 has been observed to cross-link to RNA residues within each critical sequence component of the intron: The consensus sequences that define the 5ЈSS, the branch-point (BP), and the 3Ј splice site (3ЈSS) (MacMillan et

show abstract

Removal of an intron with unique 3′ branch site creates an amino-terminal protein sequence directing the scERV1 gene product to mitochondria

Lisowsky

1996

Yeast

View full text Add to dashboard Cite

The yeast scERV1 gene is of special interest because it has a dual function in mitochondrial biogenesis and in the regulation of the cell cycle. The recent discovery that the yeast scERV1 gene has a structural and functional human homologue initiated a detailed comparison of the genes and their structures. In addition the homologous ALR (augmenter of liver regeneration) genes from rat and mouse have just been identified and it has been found that the mammalian proteins have a specific function in liver regeneration and in spermatogenesis. It now turns out that the organization of the 5′ regions of these genes is much more complicated than expected. The latest research has discovered an additional intron in the 5′ region of the mouse gene and possible amino‐terminal extensions of the reading frames. In this work, reinvestigation of the 5′ region of the yeast gene identifies a putative intron with an unusual 3′ branch site. It is shown that a small intron of 83 nucleotides is present in this genomic region. Analysis of cDNA clones demonstrates that the intron is correctly removed from the messenger RNA and that therefore the unusual 3′ branch site is probably functional. Furthermore, studies with antibodies directed against recombinant scERV1 protein demonstrate that the gene product is associated with mitochondria, in agreement with its involvement in mitochondrial biogenesis. Complementation experiments with mutants and different 5′ deletions of the gene identify the corresponding promotor for transcription and the start codon for translation.

show abstract

The interaction between the first and last intron nucleotides in the second step of pre-mRNA splicing is independent of other conserved intron nucleotides

Cited by 20 publications

References 41 publications

A mutational analysis of U12-dependent splice site dinucleotides

A mutational analysis of U12-dependent splice site dinucleotides

Allele-specific genetic interactions between Prp8 and RNA active site residues suggest a function for Prp8 at the catalytic core of the spliceosome

Removal of an intron with unique 3′ branch site creates an amino-terminal protein sequence directing the scERV1 gene product to mitochondria

Contact Info

Product

Resources

About