scholarly journals Tissue-specific and developmentally regulated alternative splicing in mouse skeletal muscle ryanodine receptor mRNA

1995 ◽  
Vol 305 (2) ◽  
pp. 373-378 ◽  
Author(s):  
A Futatsugi ◽  
G Kuwajima ◽  
K Mikoshiba
Keyword(s):  
Skeletal Muscle ◽  
Alternative Splicing ◽  
Ryanodine Receptor ◽  
Splice Variants ◽  
Amino Acid Sequences ◽  
Pcr Analysis ◽  
Developmentally Regulated ◽  
Tissue Specific ◽  
Mouse Skeletal Muscle ◽  
Splicing Patterns

The ryanodine receptor is a channel for Ca2+ release from intracellular stores. By PCR analysis, we identified two alternatively spliced regions in mRNA of the mouse skeletal muscle ryanodine receptor (sRyR). The splice variants were characterized by the presence or absence of 15 bp (ASI) and 18 bp (ASII) exons. The exclusion of these exons results in the absence of the regions corresponding to Ala3481-Gln3485 and Val3865-Asn3870, respectively, of rabbit sRyR; these amino acid sequences exist in the modulatory region, where sites for phosphorylation and binding of Ca2+, calmodulin and ATP are postulated to be. We also detected sRyR in brain and heart as well as in skeletal muscle, and the splicing patterns were found to be tissue-specific. Only the ASII-lacking isoform was detected in heart, whereas in other tissues the ASII-containing isoform was predominant. The splicing patterns were also found to change during development. In skeletal muscle, the ASI-containing isoform increased gradually from embryo to adult. The ASII-lacking isoform abruptly increased upon birth, but the ASII-containing isoform increased steadily afterwards. In cerebrum, the ratio of the ASII-containing isoform to the ASII-lacking one increased abruptly during embryonic days 14 and 18. These findings suggest that the alternative splicing of ASI and ASII, by affecting the modulatory region, generates functionally different sRyR isoforms in a tissue-specific and developmentally regulated manner.

scholarly journals Primary structure of rat plasma membrane Ca2+-ATPase isoform 4 and analysis of alternative splicing patterns at splice site A

1995 ◽  
Vol 306 (3) ◽  
pp. 779-785 ◽  
Author(s):  
T P Keeton ◽  
G E Shull
Keyword(s):  
Plasma Membrane ◽  
Alternative Splicing ◽  
Splice Site ◽  
Primary Structure ◽  
Sequence Divergence ◽  
Rat Plasma ◽  
Pcr Analysis ◽  
Rat Tissues ◽  
Mrna Tissue Distribution ◽  
Splicing Patterns

We have determined the primary structure of the rat plasma membrane Ca(2+)-ATPase isoform 4 (PMCA4), and have analysed its mRNA tissue distribution and alternative splicing patterns at splice site A. Rat PMCA4 (rPMCA4) genomic clones were isolated and used to determine the coding sequences and intron/exon organization of the 5′-end of the gene, and the remaining coding sequence was determined from PCR-amplified cDNA fragments. Pairwise comparisons reveal that the amino acid sequence of rPMCA4 has diverged substantially from those of rPMCA isoforms 1, 2 and 3 (73-76% identity) and from that of human PMCA4 (87%). Despite the high degree of sequence divergence between the two species, comparisons of intron and untranslated mRNA sequences with the corresponding human sequences confirm the identity of this rat isoform as PMCA4. Northern blot studies demonstrate that the PMCA4 mRNA is expressed in all rat tissues examined except liver, with the highest levels in uterus and stomach. A combination of PCR analysis of alternative splicing patterns and sequence analysis of the gene demonstrate that a 36 nt exon at site A is included in PMCA4 mRNAs of most tissues but is largely excluded in heart and testis. Alternative splicing of both the 36 nt exon and a previously characterized 175 nt exon at splice site C, each of which can be either included or excluded in a highly tissue-specific manner, leads to the production of four different PMCA4 variants ranging in size from 1157 to 1203 amino acids.

Tissue specificity and alternative splicing of the Na+/Ca2+ exchanger isoforms NCX1, NCX2, and NCX3 in rat

1997 ◽  
Vol 272 (4) ◽  
pp. C1250-C1261 ◽  
Author(s):  
B. D. Quednau ◽  
D. A. Nicoll ◽  
K. D. Philipson
Keyword(s):  
Skeletal Muscle ◽  
Polymerase Chain Reaction ◽  
Alternative Splicing ◽  
Reverse Transcriptase ◽  
Splice Variants ◽  
Variable Region ◽  
Intracellular Loop ◽  
Chain Reaction ◽  
Splicing Isoforms ◽  
Polymerase Chain

The gene coding for the Na+/Ca2+ exchanger NCX1 is characterized by a cluster of six exons (A, B, C, D, E, and F) coding for a variable region in the COOH terminus of the large intracellular loop of the protein. Alternative splicing of these exons generates multiple tissue-specific variants of NCX1. Using reverse transcriptase-polymerase chain reaction, we analyzed eight previously described and four new splicing isoforms of NCX1 in a wide variety of tissues and cells. Exons A and B are mutually exclusive, as shown in earlier studies, and splicing isoforms containing exon A are preferentially expressed in heart, brain, and skeletal muscle, whereas splicing variants with exon B are found in all rat tissues except heart. The second and third isoforms of the Na+/Ca2+ exchanger, NCX2 and NCX3, show a deletion of 37 amino acids in the intracellular loop corresponding to parts of the variable region of NCX1. We identified three splicing isoforms of NCX3 in brain and skeletal muscle by reverse transcriptase-polymerase chain reaction. These splice variants are generated by including either of two alternative exons equivalent to the NCX1 exon A or B and by including or excluding a sequence equivalent to the NCX1 exon C. We did not detect any alternative splicing of NCX2. We examined selected tissues from neonatal and adult rats and found developmental regulation for NCX1 and NCX3 splicing isoforms in skeletal muscle. Specific isoform patterns were also detected for NCX1 and NCX3 in cultured cortical neurons, astrocytes, and oligodendrocytes. We suggest a new terminology to distinguish the different splice variants of individual NCX isoforms.

scholarly journals Insulin receptor substrate 1 binds two novel splice variants of the regulatory subunit of phosphatidylinositol 3-kinase in muscle and brain.

1996 ◽  
Vol 16 (5) ◽  
pp. 2195-2203 ◽  
Author(s):  
D A Antonetti ◽  
P Algenstaedt ◽  
C R Kahn
Keyword(s):  
Skeletal Muscle ◽  
Insulin Receptor ◽  
Tissue Distribution ◽  
Cardiac Muscle ◽  
Splice Variants ◽  
Regulatory Subunit ◽  
Insulin Receptor Substrate ◽  
Insulin Receptor Substrate 1 ◽  
Tissue Specific ◽  
Sh2 Domains

We have identified two novel alternatively spliced forms of the p85alpha regulatory subunit of phosphatidylinositol (PI) 3-kinase by expression screening of a human skeletal muscle library with phosphorylated baculovirus- produced human insulin receptor substrate 1. One form is identical to p85alpha throughout the region which encodes both Src homology 2 (SH2) domains and the inter-SH2 domain/p110 binding region but diverges in sequence from p85alpha on the 5' side of nucleotide 953, where the entire break point cluster gene and SH3 regions are replaced by a unique 34-amino-acid N terminus. This form has an estimated molecular mass of approximately 53 kDa and has been termed p85/AS53. The second form is identical to p85 and p85/AS53 except for a 24-nucleotide insert between the SH2 domains that results in a replacement of aspartic acid 605 with nine amino acids, adding two potential serine phosphorylation sites in the vicinity of the known serine autophosphorylation site (Ser-608). Northern (RNA) analyses reveal a wide tissue distribution of p85alpha, whereas p85/AS53 is dominant in skeletal muscle and brain, and the insert isoforms are restricted to cardiac muscle and skeletal muscle. Western blot (immunoblot) analyses using an anti-p85 polyclonal antibody and a specific anti-p85/AS53 antibody confirmed the tissue distribution of p85/AS53 protein and indicate a approximately 7-fold higher expression of p85/AS53 protein than of p85 in skeletal muscle. Both p85 and p85/AS53 bind to p110 in coprecipitation experiments, but p85alpha itself appears to have preferential binding to insulin receptor substrate 1 following insulin stimulation. These data indicate that the gene for the p85alpha regulatory subunit of PI 3-kinase can undergo tissue-specific alternative splicing. Two novel splice variants of the regulatory subunit of PI 3-kinase are present in skeletal muscle, cardiac muscle, and brain; these variants may have important functional differences in activity and may play a role in tissue-specific signals such as insulin-stimulated glucose transport or control of neurotransmitter secretion or action.

scholarly journals Spontaneous and voltage-activated Ca2+release in adult mouse skeletal muscle fibres expressing the type 3 ryanodine receptor

2008 ◽  
Vol 586 (2) ◽  
pp. 441-457 ◽  
Author(s):  
Claude Legrand ◽  
Emiliana Giacomello ◽  
Christine Berthier ◽  
Bruno Allard ◽  
Vincenzo Sorrentino ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Ryanodine Receptor ◽  
Adult Mouse ◽  
Muscle Fibres ◽  
Mouse Skeletal Muscle ◽  
Skeletal Muscle Fibres ◽  
Type 3

scholarly journals Calcium current modulation by the γ1 subunit depends on alternative splicing of CaV1.1

2021 ◽  
Author(s):  
Yousra El El Ghaleb ◽  
Nadine J. Ortner ◽  
Wilfried Posch ◽  
Monica L. Fernandez-Quintero ◽  
Wietske E. Tuinte ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Current Density ◽  
Voltage Dependence ◽  
Splice Variants ◽  
Calcium Currents ◽  
Developmentally Regulated ◽  
The Difference ◽  
Ion Conducting ◽  
And Shifts ◽  
Oppositely Charged

The skeletal muscle voltage-gated calcium channel (CaV1.1) primarily functions as voltage sensor for excitation-contraction coupling. Conversely, its ion-conducting function is modulated by multiple mechanisms within the pore-forming α1S subunit and the auxiliary α2δ-1 and γ1 subunits. Particularly, developmentally regulated alternative splicing of exon 29, which inserts 19 amino acids in the extracellular IVS3-S4 loop of CaV1.1a, greatly reduces the current density and shifts the voltage-dependence of activation to positive potentials outside the physiological range. We generated a new HEK293-cell line stably expressing α2δ-1, β3, and STAC3. When the adult (CaV1.1a) and the embryonic (CaV1.1e) splice variants were expressed in these cells, the difference in the voltage-dependence of activation observed in muscle cells was reproduced, but not the reduced current density of CaV1.1a. Only when we further co-expressed the γ1 subunit, the current density of CaV1.1a, but not of CaV1.1e, was reduced by >50 %. In addition, γ1 caused a shift of the voltage-dependence of inactivation to negative voltages in both variants. Thus, the current-reducing effect of γ1, but not its effect on inactivation, is specifically dependent on the inclusion of exon 29 in CaV1.1a. Molecular structure modeling revealed several direct ionic interactions between oppositely charged residues in the IVS3-S4 loop and the γ1 subunit. However, substitution of these residues by alanine, individually or in combination, did not abolish the γ1-dependent reduction of current density, suggesting that structural rearrangements of CaV1.1a induced by inclusion of exon 29 allosterically empower the γ1 subunit to exert its inhibitory action on CaV1.1 calcium currents.

scholarly journals Ankyrin-G in skeletal muscle: Tissue-specific alternative splicing contributes to the complexity of the sarcolemmal cytoskeleton☆

2005 ◽  
Vol 309 (1) ◽  
pp. 86-98 ◽  
Author(s):  
Alexander A. Hopitzan ◽  
Anthony J. Baines ◽  
Marie-Aline Ludosky ◽  
Michel Recouvreur ◽  
Ekaterini Kordeli
Keyword(s):  
Skeletal Muscle ◽  
Alternative Splicing ◽  
Muscle Tissue ◽  
Skeletal Muscle Tissue ◽  
Tissue Specific ◽  
Ankyrin G ◽  
Specific Alternative

Tissue-specific alternative splicing of mouse brain type ryanodine receptor/calcium release channel mRNA

FEBS Letters ◽  
1996 ◽  
Vol 395 (2-3) ◽  
pp. 123-126 ◽  
Author(s):  
Ryosuke Miyatake ◽  
Aizo Furukawa ◽  
Masayuki Matsushita ◽  
Kazuhiko Iwahashi ◽  
Kazuhiko Nakamura ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Ryanodine Receptor ◽  
Mouse Brain ◽  
Calcium Release ◽  
Calcium Release Channel ◽  
Release Channel ◽  
Tissue Specific ◽  
Specific Alternative

Alternative Splicing of Rat Tropoelastin mRNA is Tissue-Specific and Developmentally Regulated

Matrix ◽  
1991 ◽  
Vol 11 (5) ◽  
pp. 359-366 ◽  
Author(s):  
Ruth A. Heim ◽  
Richard A. Pierce ◽  
Susan B. Deak ◽  
David J. Riley ◽  
Charles D. Boyd ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Developmentally Regulated ◽  
Tissue Specific
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