Control of calcitonin/calcitonin gene-related peptide pre-mRNA processing by constitutive intron and exon elements.

Yeakley, Joanne M.; Hedjran, Farah; Morfin, J P; Merillat, N; Rosenfeld, M G; Emeson, Ronald B.

doi:10.1128/mcb.13.10.5999

Cited by 66 publications

(57 citation statements)

References 77 publications

(91 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The sequence motif involved in nonsense-mediated mRNA decay is another example of a short RNA repeat involved in posttranscriptional regulation. Short RNA sequences that modulate both mRNA turnover and splicing have been identified (10,27,33,34,36,37,63,68,73,74,77). As with the results presented here, sequences flanking these motifs can influence their activity.…”

Section: Discussionsupporting

confidence: 77%

Identification and Characterization of a Sequence Motif Involved in Nonsense-Mediated mRNA Decay

Zhang

Ruiz-Echevarría²,

Quan

et al. 1995

Molecular and Cellular Biology

123

View full text Add to dashboard Cite

In both prokaryotes and eukaryotes, nonsense mutations in a gene can enhance the decay rate or reduce the abundance of the mRNA transcribed from that gene, and we call this process nonsense-mediated mRNA decay. We have been investigating the cis-acting sequences involved in this decay pathway. Previous experiments have demonstrated that, in addition to a nonsense codon, specific sequences 3 of a nonsense mutation, which have been defined as downstream elements, are required for mRNA destabilization. The results presented here identify a sequence motif (TGYYGATGYYYYY, where Y stands for either T or C) that can predict regions in genes that, when positioned 3 of a nonsense codon, promote rapid decay of its mRNA. Sequences harboring two copies of the motif from five regions in the PGK1, ADE3, and HIS4 genes were able to function as downstream elements. In addition, four copies of this motif can function as an independent downstream element. The sequences flanking the motif played a more significant role in modulating its activity when fewer copies of the sequence motif were present. Our results indicate the sequences 5 of the motif can modulate its activity by maintaining a certain distance between the sequence motif and the termination codon. We also suggest that the sequences 3 of the motif modulate the activity of the downstream element by forming RNA secondary structures. Consistent with this view, a stem-loop structure positioned 3 of the sequence motif can enhance the activity of the downstream element. This sequence motif is one of the few elements that have been identified that can predict regions in genes that can be involved in mRNA turnover. The role of these sequences in mRNA decay is discussed.The rates at which mRNAs decay play an important role in regulating gene expression. mRNA half-lives can vary from each other by as much as 100-fold (28,55,59,62,64), and this variation determines, in part, the cytoplasmic abundance of these RNAs. In addition, the decay rate of an mRNA can be modulated, and its half-life can vary depending on the cell type or the environmental situation of the cell (e.g., hormones, stress, cell cycle, etc. [3,55,62]). The goal of our work is to understand the cellular mechanisms that determine the stability of mRNAs.We are studying mRNA decay in the yeast Saccharomyces cerevisiae. Our results, as well as work from other laboratories, strongly indicate that the processes of mRNA turnover and translation are intimately linked and that understanding of this relationship is critical in elucidating the mechanism of mRNA decay (2, 5, 9, 11, 15, 21, 23, 29-31, 41, 53, 55-59, 76). The role of translation in determination of mRNA decay rates is direct, and, for certain instability elements identified in protein coding regions and 3Ј untranslated regions (3Ј-UTRs), translation of the protein coding region must occur in order for them to function (2,11,15,21,23,29,41,53,60,66,76).One clear example of the relationship between translation and mRNA decay is the observation that nonsense mutati...

show abstract

Section: Discussionsupporting

confidence: 77%

Identification and Characterization of a Sequence Motif Involved in Nonsense-Mediated mRNA Decay

Zhang

Ruiz-Echevarría²,

Quan

et al. 1995

Molecular and Cellular Biology

123

View full text Add to dashboard Cite

show abstract

“…Support for our hypothesis is provided by the identification of several sequence elements, including three exon 4 sequence motifs, a sequence upstream of the exon 4, and a sequence downstream of exon 4 (Emeson et al 1989, Cote et al 1992, Roesser et al 1993, van Oers et al 1994, Yeakley et al 1993. We have focused on a sequence element located downstream of exon 4, because it has several interesting features.…”

mentioning

confidence: 68%

Alternative RNA processing--its role in regulating expression of calcitonin/calcitonin gene-related peptide

Lou

Gagel²

1998

Journal of Endocrinology

View full text Add to dashboard Cite

The calcitonin/calcitonin gene-related peptide (CT/ CGRP) gene is one of the earliest studied examples of alternative RNA processing. The regulatory mechanisms controlling this event are poorly understood. We have identified and characterized an intron element residing in intron 4 of the human CT/CGRP gene. This intron element functions to enhance polyadenylation of an embedded alternative 3 -terminal exon within the CT/ CGRP gene and is potentially involved in tissue-specific regulation of CT/CGRP RNA processing. (1998) 156, 401-405 In eukaryotic nuclei, every pre-messenger RNA (premRNA) undergoes post-transcriptional processing to generate mature messenger RNA (mRNA), which is exported to the cytoplasm for translation. Posttranscriptional modification is a complex process, including capping at the 5 end, polyadenylation at the 3 end, and splicing or removal of intervening sequences (introns); splicing results in the precise ligation of coding-sequencecontaining exons. Transcription and RNA processing are tightly coupled (Steinmetz 1997): there is compelling evidence that interaction of RNA processing factors with the newly transcribed RNA occurs during transcription (Daneholt 1997). Capping and polyadenylation occur concomitantly with transcription, but splicing occurs during or after transcription (Beyer & Osheim 1988). Journal of EndocrinologyErrors of splicing and polyadenylation can cause the production of non-functional proteins, and form the molecular basis of a large number of diseases, including endocrine diseases (Harteveld et al. 1994, Nakai & Sakamoto 1994. For example, in one case of hereditary vitamin D rickets, a G to C point mutation at the +5 position of the 5 donor splice site of intron 4 caused exon 4 skipping, which leads to the production of a truncated protein lacking the hormone-binding domain, through a frame-shift in translation (Hawa et al. 1996). In another example, in patients from several families in which familial persisitent hyperinsulinemic hypoglycemia of infancy occurred, a mutation at the 3 acceptor splice site was identified to cause altered splicing with cryptic 3 splice sites leading to a defective sulfonylurea receptor (Thomas et al. 1995). In a third example, several common mutations have been mapped to CYP21B in steroid 21-hydroxylase deficiency, the leading cause of impaired cortisol synthesis in congenital adrenal hyperplasia. One of the mutations, located in intron 2, causes abnormal splicing (Owerbach et al. 1990).Splicing is mediated by several families of trans-acting factors that interact with specific RNA sequences (Moore et al. 1993). These include 5 and 3 (polypyrimidine-tract and branchpoint sequence) splice site sequences that define exon-intron boundaries ( Fig. 1; Moore et al. 1993). Several new classes of exon and intron elements have now been identified that interact with the rapidly growing family of RNA binding proteins (Lou et al. 1995). Two prominent members of this family include small nuclear RNA (snRNA)-containing small nuclear ribonuc...

show abstract

“…Changing the noncanonical branchpoint upstream of exon 4 to the preferred A allowed usage of the exon 4 splice acceptor in nuclear extract from calcitonin-producing cells, but not in nuclear extract from CGRP-producing cells (Cote et al 1991). Deletion of the exon 5 splice acceptor was shown to abolish CGRP splicing in CGRP-producing cells, but did not increase the level of calcitonin splicing (Yeakley et al 1993). 6-9).…”

Section: Discussionmentioning

confidence: 99%

“…1A). Several elements have been identified in and around exon 4 that control exon 4 inclusion into mRNA and use of the calcitonin-specific polyadenylation site in intron 4 (Leff et al 1987;Adema et al , 1990Roesser et al 1993;Yeakley et al 1993;van Oers et al 1994;Lou et al 1995Lou et al , 1998Lou et al , 1999Zandberg et al 1995;Coleman and Roesser 1998). To determine whether sequences in or around exon 5 also play a role in the regulation of calcitonin/CGRP mRNA processing, RNA from minigenes containing exon 5 were tested in in vitro splicing reactions.…”

Section: Splicing Of Calcitonin Exon 5 In Vitromentioning

confidence: 99%

Both U2 snRNA and U12 snRNA are required for accurate splicing of exon 5 of the rat calcitonin/CGRP gene

Roesser

2004

RNA

View full text Add to dashboard Cite

Two classes of spliceosome are present in eukaryotic cells. Most introns in nuclear pre-mRNAs are removed by a spliceosome that requires U1, U2, U4, U5, and U6 small nuclear ribonucleoprotein particles (snRNPs). A minor class of introns are removed by a spliceosome containing U11, U12, U5, U4atac, and U6 atac snRNPs. We describe experiments that demonstrate that splicing of exon 5 of the rat calcitonin/CGRP gene requires both U2 snRNA and U12 snRNA. In vitro, splicing to calcitonin/ CGRP exon 5 RNA was dependent on U2 snRNA, as preincubation of nuclear extract with an oligonucleotide complementary to U2 snRNA abolished exon 5 splicing. Addition of an oligonucleotide complementary to U12 snRNA increased splicing at a cryptic splice site in exon 5 from <5% to 50% of total spliced RNA. Point mutations in a candidate U12 branch sequence in calcitonin/CGRP intron 4, predicted to decrease U12-pre-mRNA base-pairing, also significantly increased cryptic splicing in vitro. Calcitonin/CGRP genes containing base changes disrupting the U12 branch sequence expressed significantly decreased CGRP mRNA levels when expressed in cultured cells. Coexpression of U12 snRNAs containing base changes predicted to restore U12-pre-mRNA base pairing increased CGRP mRNA synthesis to the level of the wild-type gene. These observations indicate that accurate, efficient splicing of calcitonin/CGRP exon 5 is dependent upon both U2 and U12 snRNAs.

show abstract

Control of calcitonin/calcitonin gene-related peptide pre-mRNA processing by constitutive intron and exon elements.

Cited by 66 publications

References 77 publications

Identification and Characterization of a Sequence Motif Involved in Nonsense-Mediated mRNA Decay

Identification and Characterization of a Sequence Motif Involved in Nonsense-Mediated mRNA Decay

Alternative RNA processing--its role in regulating expression of calcitonin/calcitonin gene-related peptide

Both U2 snRNA and U12 snRNA are required for accurate splicing of exon 5 of the rat calcitonin/CGRP gene

Contact Info

Product

Resources

About