Two different sequence elements within exon 4 are necessary for calcitonin-specific splicing of the human calcitonin/calcitonin gene-related peptide I pre-mRNA

1994 ◽  
Vol 14 (2) ◽  
pp. 951-960
Author(s):  
C C van Oers ◽  
G J Adema ◽  
H Zandberg ◽  
T C Moen ◽  
P D Baas
Keyword(s):  
Calcitonin Gene Related Peptide ◽  
Splice Acceptor Site ◽  
Acceptor Site ◽  
Sequence Element ◽  
Tissue Specific ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Sequence Elements ◽  
Exon 4 ◽  
I Gene

The calcitonin (CT)/calcitonin gene-related peptide I (CGRP-I) gene (CALC-I gene) is subject to alternative tissue-specific processing of its primary transcript. CT mRNA is the predominant mRNA produced in thyroid C cells, whereas CT gene-related peptide I mRNA is the main product in neurons of the central and peripheral nervous systems. The CT-specific exon 4 is surrounded by weak processing sites. In this study we have investigated whether exon 4 sequences are involved in the tissue-specific selection of the exon 4 splice acceptor site. The results indicate that two separate elements, termed A and B, in the 5' part of exon 4 are required for production of CT-specific RNA. These sequences are located between nucleotides 67 and 88 (A) and nucleotides 117 and 146 (B) relative to the 5' end of exon 4. Variation of the distance between these sequence elements and the 3' splice site of exon 4 does not change the processing choice. These sequence elements are functionally equivalent. CT-specific splicing requires the presence of both sequence A and B or duplicates of either sequence element in exon 4. The effect of these sequences on the RNA processing choice is overruled by mutation of the CT-specific uridine branch acceptor nucleotide into a commonly preferred adenosine residue.

scholarly journals Two different sequence elements within exon 4 are necessary for calcitonin-specific splicing of the human calcitonin/calcitonin gene-related peptide I pre-mRNA.

1994 ◽  
Vol 14 (2) ◽  
pp. 951-960 ◽  
Author(s):  
C C van Oers ◽  
G J Adema ◽  
H Zandberg ◽  
T C Moen ◽  
P D Baas
Keyword(s):  
Calcitonin Gene Related Peptide ◽  
Splice Acceptor Site ◽  
Acceptor Site ◽  
Sequence Element ◽  
Tissue Specific ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Sequence Elements ◽  
Exon 4 ◽  
I Gene

The calcitonin (CT)/calcitonin gene-related peptide I (CGRP-I) gene (CALC-I gene) is subject to alternative tissue-specific processing of its primary transcript. CT mRNA is the predominant mRNA produced in thyroid C cells, whereas CT gene-related peptide I mRNA is the main product in neurons of the central and peripheral nervous systems. The CT-specific exon 4 is surrounded by weak processing sites. In this study we have investigated whether exon 4 sequences are involved in the tissue-specific selection of the exon 4 splice acceptor site. The results indicate that two separate elements, termed A and B, in the 5' part of exon 4 are required for production of CT-specific RNA. These sequences are located between nucleotides 67 and 88 (A) and nucleotides 117 and 146 (B) relative to the 5' end of exon 4. Variation of the distance between these sequence elements and the 3' splice site of exon 4 does not change the processing choice. These sequence elements are functionally equivalent. CT-specific splicing requires the presence of both sequence A and B or duplicates of either sequence element in exon 4. The effect of these sequences on the RNA processing choice is overruled by mutation of the CT-specific uridine branch acceptor nucleotide into a commonly preferred adenosine residue.

Calcitonin/calcitonin gene-related peptide transcription unit: tissue-specific expression involves selective use of alternative polyadenylation sites

1984 ◽  
Vol 4 (10) ◽  
pp. 2151-2160
Author(s):  
S G Amara ◽  
R M Evans ◽  
M G Rosenfeld
Keyword(s):  
Regulation Of Gene Expression ◽  
Alternative Polyadenylation ◽  
Calcitonin Gene Related Peptide ◽  
Transcription Unit ◽  
Specific Expression ◽  
Tissue Specific ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Polyadenylation Sites ◽  
Specific Regulation

Different 3' coding exons in the rat calcitonin gene are used to generate distinct mRNAs encoding either the hormone calcitonin in thyroidal C-cells or a new neuropeptide referred to as calcitonin gene-related peptide in neuronal tissue, indicating the RNA processing regulation is one strategy used in tissue-specific regulation of gene expression in the brain. Although the two mRNAs use the same transcriptional initiation site and have identical 5' terminal sequences, their 3' termini are distinct. The polyadenylation sites for calcitonin and calcitonin gene-related peptide mRNAs are located at the end of the exons 4 and 6, respectively. Termination of transcription after the calcitonin exon does not dictate the production of calcitonin mRNA, because transcription proceeds through both calcitonin and calcitonin gene-related peptide exons irrespective of which mRNA is ultimately produced. In isolated nuclei, both polyadenylation sites appear to be utilized; however, the proximal (calcitonin) site is preferentially used in nuclei from tissues producing calcitonin mRNA. These data suggest that the mechanism dictating production of each mRNA involves the selective use of alternative polyadenylation sites.

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

1993 ◽  
Vol 13 (10) ◽  
pp. 5999-6011
Author(s):  
J M Yeakley ◽  
F Hedjran ◽  
J P Morfin ◽  
N Merillat ◽  
M G Rosenfeld ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Calcitonin Gene Related Peptide ◽  
Regulatory Sequences ◽  
Primary Transcript ◽  
Tissue Specific ◽  
Related Peptide ◽  
Thyroid C Cells ◽  
Calcitonin Gene ◽  
Constitutive Splicing ◽  
Specific Regulation

The calcitonin/calcitonin gene-related peptide (CGRP) primary transcript is alternatively spliced in thyroid C cells and neurons, resulting in the tissue-specific production of calcitonin and CGRP mRNAs. Analyses of mutated calcitonin/CGRP transcription units in permanently transfected cell lines have indicated that alternative splicing is regulated by a differential capacity to utilize the calcitonin-specific splice acceptor. The analysis of an extensive series of mutations suggests that tissue-specific regulation of calcitonin mRNA production does not depend on the presence of a single, unique cis-active element but instead appears to be a consequence of suboptimal constitutive splicing signals. While only those mutations that altered constitutive splicing signals affected splice choices, the action of multiple regulatory sequences cannot be formally excluded. Further, we have identified a 13-nucleotide purine-rich element from a constitutive exon that, when placed in exon 4, entirely switches splice site usage in CGRP-producing cells. These data suggest that specific exon recruitment sequences, in combination with other constitutive elements, serve an important function in exon recognition. These results are consistent with the hypothesis that tissue-specific alternative splicing of the calcitonin/CGRP primary transcript is mediated by cell-specific differences in components of the constitutive splicing machinery.

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

1998 ◽  
Vol 156 (3) ◽  
pp. 401-405 ◽  
Author(s):  
H Lou ◽  
RF Gagel
Keyword(s):  
Rna Processing ◽  
Calcitonin Gene Related Peptide ◽  
Regulatory Mechanisms ◽  
Terminal Exon ◽  
Tissue Specific ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Specific Regulation ◽  
Intron 4

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.

scholarly journals Calcitonin/calcitonin gene-related peptide transcription unit: tissue-specific expression involves selective use of alternative polyadenylation sites.

1984 ◽  
Vol 4 (10) ◽  
pp. 2151-2160 ◽  
Author(s):  
S G Amara ◽  
R M Evans ◽  
M G Rosenfeld
Keyword(s):  
Regulation Of Gene Expression ◽  
Alternative Polyadenylation ◽  
Calcitonin Gene Related Peptide ◽  
Transcription Unit ◽  
Specific Expression ◽  
Tissue Specific ◽  
Related Peptide ◽  
Calcitonin Gene ◽  
Polyadenylation Sites ◽  
Specific Regulation

Different 3' coding exons in the rat calcitonin gene are used to generate distinct mRNAs encoding either the hormone calcitonin in thyroidal C-cells or a new neuropeptide referred to as calcitonin gene-related peptide in neuronal tissue, indicating the RNA processing regulation is one strategy used in tissue-specific regulation of gene expression in the brain. Although the two mRNAs use the same transcriptional initiation site and have identical 5' terminal sequences, their 3' termini are distinct. The polyadenylation sites for calcitonin and calcitonin gene-related peptide mRNAs are located at the end of the exons 4 and 6, respectively. Termination of transcription after the calcitonin exon does not dictate the production of calcitonin mRNA, because transcription proceeds through both calcitonin and calcitonin gene-related peptide exons irrespective of which mRNA is ultimately produced. In isolated nuclei, both polyadenylation sites appear to be utilized; however, the proximal (calcitonin) site is preferentially used in nuclei from tissues producing calcitonin mRNA. These data suggest that the mechanism dictating production of each mRNA involves the selective use of alternative polyadenylation sites.

scholarly journals Role for Fox-1/Fox-2 in Mediating the Neuronal Pathway of Calcitonin/Calcitonin Gene-Related Peptide Alternative RNA Processing

2006 ◽  
Vol 27 (3) ◽  
pp. 830-841 ◽  
Author(s):  
Hua-Lin Zhou ◽  
Andrew P. Baraniak ◽  
Hua Lou
Keyword(s):  
Splice Site ◽  
Critical Role ◽  
Regulatory Elements ◽  
Calcitonin Gene Related Peptide ◽  
Related Peptide ◽  
Alternative Processing ◽  
Neuronal Pathway ◽  
Calcitonin Gene ◽  
Splicing Regulatory Elements ◽  
Exon 4

ABSTRACT Although multiple regulatory elements and protein factors are known to regulate the non-neuronal pathway of alternative processing of the calcitonin/calcitonin gene-related peptide (CGRP) pre-mRNA, the mechanisms controlling the neuron-specific pathway have remained elusive. Here we report the identification of Fox-1 and Fox-2 proteins as novel regulators that mediate the neuron-specific splicing pattern. Fox-1 and Fox-2 proteins function to repress exon 4 inclusion, and this effect depends on two UGCAUG elements surrounding the 3′ splice site of the calcitonin-specific exon 4. In neuron-like cells, mutation of a subset of UGCAUG elements promotes the non-neuronal pattern in which exon 4 is included. In HeLa cells, overexpression of Fox-1 or Fox-2 protein decreases exon 4 inclusion. Fox-1 and Fox-2 proteins interact with the UGCAUG elements specifically and regulate splicing by blocking U2AF65 binding to the 3′ splice site upstream of exon 4. We further investigated the inter-relationship between the UGCAUG silencer elements and the previously identified intronic and exonic splicing regulatory elements and found that exon 4 is regulated by an intricate balance of positive and negative regulation. These results define a critical role for Fox-1 and Fox-2 proteins in exon 4 inclusion of calcitonin/CGRP pre-mRNA and establish a regulatory network that controls the fate of exon 4.

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

1993 ◽  
Vol 13 (10) ◽  
pp. 5999-6011 ◽  
Author(s):  
J M Yeakley ◽  
F Hedjran ◽  
J P Morfin ◽  
N Merillat ◽  
M G Rosenfeld ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Calcitonin Gene Related Peptide ◽  
Regulatory Sequences ◽  
Primary Transcript ◽  
Tissue Specific ◽  
Related Peptide ◽  
Thyroid C Cells ◽  
Calcitonin Gene ◽  
Constitutive Splicing ◽  
Specific Regulation

The calcitonin/calcitonin gene-related peptide (CGRP) primary transcript is alternatively spliced in thyroid C cells and neurons, resulting in the tissue-specific production of calcitonin and CGRP mRNAs. Analyses of mutated calcitonin/CGRP transcription units in permanently transfected cell lines have indicated that alternative splicing is regulated by a differential capacity to utilize the calcitonin-specific splice acceptor. The analysis of an extensive series of mutations suggests that tissue-specific regulation of calcitonin mRNA production does not depend on the presence of a single, unique cis-active element but instead appears to be a consequence of suboptimal constitutive splicing signals. While only those mutations that altered constitutive splicing signals affected splice choices, the action of multiple regulatory sequences cannot be formally excluded. Further, we have identified a 13-nucleotide purine-rich element from a constitutive exon that, when placed in exon 4, entirely switches splice site usage in CGRP-producing cells. These data suggest that specific exon recruitment sequences, in combination with other constitutive elements, serve an important function in exon recognition. These results are consistent with the hypothesis that tissue-specific alternative splicing of the calcitonin/CGRP primary transcript is mediated by cell-specific differences in components of the constitutive splicing machinery.

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