scholarly journals Sequence elements surrounding the acceptor site suppress alternative splicing of the sarco/endoplasmic reticulum Ca2+-ATPase 2 gene transcript

1997 ◽  
Vol 322 (3) ◽  
pp. 885-891 ◽  
Author(s):  
Ludo VAN DEN BOSCH ◽  
Luc MERTENS ◽  
Sofie GIJSBERS ◽  
Mark VER HEYEN ◽  
Frank WUYTACK ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Distal Part ◽  
Myogenic Differentiation ◽  
Acceptor Site ◽  
Gene Transcript ◽  
Muscle Type ◽  
Sequence Elements ◽  
Neuronal Type ◽  
Primary Gene ◽  
Broad Region

Expression of the muscle-specific 2a isoform of the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2) requires activation of an inefficient optional splice process at the 3´ end of the primary gene transcript. The sequence elements required for this regulated splice event were studied by modifying a minigene containing the 3´ end of the SERCA2 gene. An important requirement appears to be a strong muscle-specific acceptor site, as replacing it by a weak one prevented the induction of muscle-type splicing during myogenic differentiation. The induction of muscle-type splicing did not depend on positive cis-active sequences in the muscle-specific exon. On the other hand, replacement of a broad region around the acceptor site dramatically deregulated the expression pattern, as this modification strongly induced muscle-type splicing in undifferentiated muscle cells and in fibroblasts. This cis-active region is also involved in the suppression of the neuronal type of splicing. Furthermore selective replacement of the acceptor site as well as deletions or replacements in the muscle-specific exon induced muscle-type splicing to various extents in undifferentiated myogenic cells. Therefore sequence elements in the distal part of the optional intron and in the muscle-specific exon contribute to the suppression of muscle-specific SERCA2 splicing.

scholarly journals Evidence for two isoforms of the endoplasmic-reticulum Ca2+ pump in pig smooth muscle

1989 ◽  
Vol 260 (3) ◽  
pp. 757-761 ◽  
Author(s):  
J A Eggermont ◽  
F Wuytack ◽  
S De Jaegere ◽  
L Nelles ◽  
R Casteels
Keyword(s):  
Amino Acids ◽  
Endoplasmic Reticulum ◽  
Smooth Muscle ◽  
Cdna Clones ◽  
Gene Transcript ◽  
Muscle Tissues ◽  
Class 1 ◽  
Terminal Amino ◽  
Slow Twitch ◽  
Primary Gene

cDNA clones coding for the endoplasmic reticulum Ca2+-transport ATPase have been cloned from a pig smooth-muscle cDNA library. The transcripts can be divided into two classes which differ in their 3′ ends due to alternative splicing of the primary gene transcript. The class 1 cDNA encodes a protein of 997 amino acids (Mr 110,000). The class 2 protein (1042 amino acids; Mr 115,000) is completely identical to the class 1 protein except that the four C-terminal amino acids of the class 1 protein are replaced in the class 2 protein with a tail of 49 amino acids. Comparison of these sequences with other Ca2+ pump sequences reveals that the class 1 isoform corresponds to the sarcoplasmic reticulum Ca2+ pump of slow-twitch skeletal/cardiac muscle, whereas the class 2 protein corresponds to a Ca2+ pump recently detected in non-muscle tissues.

scholarly journals Alternative processing of the sarco/endoplasmic reticulum Ca2+-ATPase transcripts during muscle differentiation is a specifically regulated process

1996 ◽  
Vol 317 (3) ◽  
pp. 647-651 ◽  
Author(s):  
Ludo VAN DEN BOSCH ◽  
Luc MERTENS ◽  
Yvon CAVALOC ◽  
Martha PETERSON ◽  
Frank WUYTACK ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
B Cell ◽  
Alternative Splice ◽  
Polyadenylation Site ◽  
Cell Maturation ◽  
Gene Transcript ◽  
Alternative Processing ◽  
Splice Efficiency ◽  
B Cell Maturation ◽  
Primary Gene

Expression of the muscle-specific 2a isoform of the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2) requires activation of an otherwise inefficient splice process at the 3´-end of the primary gene transcript. We provide evidence that SERCA2 splicing is a specifically regulated process, rather than the result of an increase in general splice efficiency or a decrease in polyadenylation efficiency at the 5´-most polyadenylation site. This is indicated by the fact that changes in general splice and polyadenylation efficiency, as observed during B-cell maturation, did not affect SERCA2 splicing. Furthermore, expression and overexpression studies did not support the hypothesis that changes in the level of the alternative splice factor ASF/SF2 or other arginine and serine rich proteins are sufficient to obtain the regulation of muscle- and neuronal-specific splicing.

scholarly journals cDNA cloning, expression and chromosomal localization of the human sarco/endoplasmic reticulum Ca2+-ATPase 3 gene

1996 ◽  
Vol 318 (2) ◽  
pp. 689-699 ◽  
Author(s):  
Leonard DODE ◽  
Frank WUYTACK ◽  
Patrick F. J. KOOLS ◽  
Fouzia BABA-AISSA ◽  
Luc RAEYMAEKERS ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Nucleotide Sequence ◽  
Blot Hybridization ◽  
Cdna Probe ◽  
Human Platelets ◽  
Gene Transcript ◽  
Northern Blot Hybridization ◽  
Dna Encoding ◽  
Coding Region ◽  
Cos Cells

cDNA and genomic clones encoding human sarco/endoplasmic reticulum Ca2+-ATPase 3 (SERCA3) were isolated. The composite nucleotide sequence of the 4.6 kb cDNA, as well as the partial structure of 25 kb of genomic DNA encoding all but the 5´ region of the gene, was determined. The nucleotide sequence coding for the last six amino acids of the pump and the 3´-untranslated region were identified within the sequence of the last exon. Northern blot hybridization analysis using cDNA probes derived from this exon detected a 4.8 kb transcript in several human tissues. Using a cDNA probe derived from the 5´-coding region an unexpected mRNA distribution pattern, consisting of two mRNA species of 4.8 and 4.0 kb, was detected in thyroid gland and bone marrow only. This is the first indication of an alternative splicing mechanism operating on the SERCA3 gene transcript, which most likely generates SERCA3 isoforms with altered C-termini. Human SERCA3 expressed in platelets and in COS cells transfected with the corresponding cDNA was detected with the previously described antibody N89 (directed against the N-terminal region of rat SERCA3) and with a new SERCA3-specific antiserum C91, directed against the extreme C-terminus of the human isoform. A monoclonal antibody PL/IM430, previously assumed to recognize SERCA3 in human platelets, does not react with the 97 kDa human SERCA3 transiently expressed in COS cells. Therefore the 97 kDa isoform detected by PL/IM430 more likely represents a novel SERCA pump, as recently suggested [Kovács, Corvazier, Papp, Magnier, Bredoux, Enyedi, Sarkadi and Enouf (1994) J. Biol. Chem. 269, 6177–6184]. Finally, by fluorescence in situ hybridization and chromosome G-banding analyses, the SERCA3 gene was assigned to human chromosome 17p13.3.

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.

RyR1-mediated moderate endoplasmic reticulum stress is required for myogenic differentiation of myoblasts

2021 ◽  
Author(s):  
Kai Qiu ◽  
Yubo Wang ◽  
Doudou Xu ◽  
Linjuan He ◽  
Xin Zhang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Cell Fate ◽  
Muscle Development ◽  
Myogenic Differentiation ◽  
C2c12 Cells ◽  
Stress Signaling ◽  
Congenital Myopathies ◽  
Myotube Formation

Abstract BackgroundCytosolic Ca2+ plays vital roles in myogenesis and muscle development. Key mutations of ryanodine receptor 1 (RyR1), a major Ca2+ release channel of endoplasmic reticulum (ER), are main causes of severe congenital myopathies. The role of RyR1 in myogenic differentiation has attracted intense research interest, however, it remains unclear. MethodsThis study employed RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts cells to explore the role of RyR1 in myogenic differentiation, myotube formation as well as the potential mechanism of RyR1-related myopathies.ResultsCytoplasmic Ca2+ concentration was significantly elevated during myogenic differentiation of both primary myogenic cells and myoblasts C2C12 cells, accompanied with a dramatic increase in RyR1 expression and resultant ER stress. Inhibition of RyR1 by siRNA-mediated silence or chemical inhibitor, dantrolene, significantly reduced cytosolic Ca2+, alleviated ER stress, and blocked multinucleated myotube formation. Moderate activation of ER stress effectively relieved myogenic differentiation stagnation induced by RyR1 suppression and demonstrated that RyR1 modulates myogenic differentiation via activation of Ca2+ -induced ER stress signaling. RyR1 knockout-induced Ca2+ leakage led to severe ER stress and excessive unfolded protein response, and drove cell fate from differentiation into apoptosis. ConclusionsTherefore, we concluded that dramatic increase in RyR1 expression is required for myogenic differentiation, and RyR1-mediated Ca2+ release leading to the activation of ER stress signaling serves a double-edged sword role during myogenic differentiation. This study contributes to a novel understanding of the role of RyR1 in muscle development and related congenital myopathies, and provides a potential target for regulation of muscle regeneration and tissue engineering.

scholarly journals Sequence and Spatial Requirements for Regulated Muscle-specific Processing of the Sarco/Endoplasmic Reticulum Ca-ATPase 2 Gene Transcript

1995 ◽  
Vol 270 (18) ◽  
pp. 11004-11011 ◽  
Author(s):  
Luc Mertens ◽  
Ludo Van Den Bosch ◽  
Hilde Verboomen ◽  
Frank Wuytack ◽  
Humbert De Smedt ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Gene Transcript

scholarly journals Stbd1 promotes glycogen clustering during endoplasmic reticulum stress and supports survival of mouse myoblasts

2020 ◽  
Vol 133 (20) ◽  
pp. jcs244855
Author(s):  
Andria A. Lytridou ◽  
Anthi Demetriadou ◽  
Melina Christou ◽  
Louiza Potamiti ◽  
Nikolas P. Mastroyiannopoulos ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Er Stress ◽  
Myogenic Differentiation ◽  
Apoptotic Pathway ◽  
Unfolded Protein ◽  
The Unfolded Protein Response ◽  
Glucose Restriction ◽  
Protein Response ◽  
Er Homeostasis ◽  
Upr Pathway

ABSTRACTImbalances in endoplasmic reticulum (ER) homeostasis provoke a condition known as ER stress and activate the unfolded protein response (UPR) pathway, an evolutionarily conserved cell survival mechanism. Here, we show that mouse myoblasts respond to UPR activation by stimulating glycogenesis and the formation of α-amylase-degradable, glycogen-containing ER structures. We demonstrate that the glycogen-binding protein Stbd1 is markedly upregulated through the PERK signalling branch of the UPR pathway and is required for the build-up of glycogen structures in response to ER stress activation. In the absence of ER stress, Stbd1 overexpression is sufficient to induce glycogen clustering but does not stimulate glycogenesis. Glycogen structures induced by ER stress are degraded under conditions of glucose restriction through a process that does not depend on autophagosome–lysosome fusion. Furthermore, we provide evidence that failure to induce glycogen clustering during ER stress is associated with enhanced activation of the apoptotic pathway. Our results reveal a so far unknown response of mouse myoblasts to ER stress and uncover a novel specific function of Stbd1 in this process, which may have physiological implications during myogenic differentiation.This article has an associated First Person interview with the first author of the paper.

Regulation of the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) 2 gene transcript in neuronal cells

1998 ◽  
Vol 55 (1) ◽  
pp. 92-100 ◽  
Author(s):  
Fawzia Baba-Aı̈ssa ◽  
Ludo Van Den Bosch ◽  
Frank Wuytack ◽  
Luc Raeymaekers ◽  
Rik Casteels
Keyword(s):  
Endoplasmic Reticulum ◽  
Neuronal Cells ◽  
Gene Transcript

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.

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