A variably spliced region in the type 1 ryanodine receptor may participate in an inter-domain interaction

Takashi Kimura, Suzy M Pace, Lan Wei, Nicole A Beard, Robert T Dirksen, Angela F. Dulhunty

    Research output: Contribution to journalArticle

    23 Citations (Scopus)

    Abstract

    The aim of the present study was to examine residues that are variably spliced in the juvenile and adult isoforms of the skeletal-muscle RyR1 (type 1 ryanodine receptor). The juvenile ASI(-) splice variant is less active than the adult ASI(+) variant and is overexpressed in patients with DM (myotonic dystrophy) [Kimura, Nakamori, Lueck, Pouliquin, Aoike, Fujimura, Dirksen, Takahashi, Dulhunty and Sakoda (2005) Hum. Mol. Genet. 14, 2189-2200]. In the present study, we explore the ASI region using synthetic peptides corresponding to rabbit RyR1 residues Thr3471-Gly3500 either containing [PASI(+)] or lacking [PASI(-)] the ASI residues. Both peptides increased [3H]ryanodine binding to rabbit RyR1s, increased Ca2+ release from sarcoplasmic reti-culum vesicles and increased single RyR1 channel activity. The peptide PASI(-) was more active in each case than PASI(+). [3H]Ryanodine binding to recombinant ASI(+)RyR1 or ASI(-)-RyR1 was enhanced more by PASI(-) than PASI(+), with the greatest increase seen when PASI(-) was added to ASI(-)RyR1. The activation of the RyR channels is consistent with the hypo-thesis that the peptides interrupt an inhibitory inter-domain inter-action and that PASI(-) is more effective at interrupting this interaction than PASI(+). We therefore suggest that the ASI(-) sequence interacts more tightly than the ASI(+) sequence with its binding partner, so that the ASI(-)RyR1 is more strongly inhibited (less active) than the ASI(+)RyR1. Thus the affinity of the binding partners in this inter-domain interaction may deter-mine the activities of the mature and juvenile isoforms of RyR1 and the stronger inhibition in the juvenile isoform may contribute to the myopathy in DM.

    Original languageEnglish
    Pages (from-to)317-324
    Number of pages8
    JournalBiochemical Journal
    Volume401
    Issue number1
    DOIs
    Publication statusPublished - 1 Jan 2007

    Fingerprint

    Ryanodine Receptor Calcium Release Channel
    Ryanodine
    Peptides
    Protein Isoforms
    Rabbits
    Viverridae
    Myotonic Dystrophy
    Muscular Diseases
    Muscle
    Skeletal Muscle
    Chemical activation

    Cite this

    Kimura, Takashi ; Pace, Suzy M ; Wei, Lan ; Beard, Nicole A ; Dirksen, Robert T ; Dulhunty, Angela F. / A variably spliced region in the type 1 ryanodine receptor may participate in an inter-domain interaction. In: Biochemical Journal. 2007 ; Vol. 401, No. 1. pp. 317-324.
    @article{5c5854e8b8cf487aad7730eee2caa857,
    title = "A variably spliced region in the type 1 ryanodine receptor may participate in an inter-domain interaction",
    abstract = "The aim of the present study was to examine residues that are variably spliced in the juvenile and adult isoforms of the skeletal-muscle RyR1 (type 1 ryanodine receptor). The juvenile ASI(-) splice variant is less active than the adult ASI(+) variant and is overexpressed in patients with DM (myotonic dystrophy) [Kimura, Nakamori, Lueck, Pouliquin, Aoike, Fujimura, Dirksen, Takahashi, Dulhunty and Sakoda (2005) Hum. Mol. Genet. 14, 2189-2200]. In the present study, we explore the ASI region using synthetic peptides corresponding to rabbit RyR1 residues Thr3471-Gly3500 either containing [PASI(+)] or lacking [PASI(-)] the ASI residues. Both peptides increased [3H]ryanodine binding to rabbit RyR1s, increased Ca2+ release from sarcoplasmic reti-culum vesicles and increased single RyR1 channel activity. The peptide PASI(-) was more active in each case than PASI(+). [3H]Ryanodine binding to recombinant ASI(+)RyR1 or ASI(-)-RyR1 was enhanced more by PASI(-) than PASI(+), with the greatest increase seen when PASI(-) was added to ASI(-)RyR1. The activation of the RyR channels is consistent with the hypo-thesis that the peptides interrupt an inhibitory inter-domain inter-action and that PASI(-) is more effective at interrupting this interaction than PASI(+). We therefore suggest that the ASI(-) sequence interacts more tightly than the ASI(+) sequence with its binding partner, so that the ASI(-)RyR1 is more strongly inhibited (less active) than the ASI(+)RyR1. Thus the affinity of the binding partners in this inter-domain interaction may deter-mine the activities of the mature and juvenile isoforms of RyR1 and the stronger inhibition in the juvenile isoform may contribute to the myopathy in DM.",
    keywords = "Alternative Splicing, Amino Acid Sequence, Animals, Exons, Genetic Variation, Kinetics, Molecular Sequence Data, Muscle, Skeletal, Rabbits, Recombinant Proteins, Ryanodine Receptor Calcium Release Channel, Sequence Alignment, Sequence Homology, Amino Acid, Transfection",
    author = "Takashi Kimura and Pace, {Suzy M} and Lan Wei and Beard, {Nicole A} and Dirksen, {Robert T} and Dulhunty, {Angela F.}",
    year = "2007",
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    A variably spliced region in the type 1 ryanodine receptor may participate in an inter-domain interaction. / Kimura, Takashi; Pace, Suzy M; Wei, Lan; Beard, Nicole A; Dirksen, Robert T; Dulhunty, Angela F.

    In: Biochemical Journal, Vol. 401, No. 1, 01.01.2007, p. 317-324.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - A variably spliced region in the type 1 ryanodine receptor may participate in an inter-domain interaction

    AU - Kimura, Takashi

    AU - Pace, Suzy M

    AU - Wei, Lan

    AU - Beard, Nicole A

    AU - Dirksen, Robert T

    AU - Dulhunty, Angela F.

    PY - 2007/1/1

    Y1 - 2007/1/1

    N2 - The aim of the present study was to examine residues that are variably spliced in the juvenile and adult isoforms of the skeletal-muscle RyR1 (type 1 ryanodine receptor). The juvenile ASI(-) splice variant is less active than the adult ASI(+) variant and is overexpressed in patients with DM (myotonic dystrophy) [Kimura, Nakamori, Lueck, Pouliquin, Aoike, Fujimura, Dirksen, Takahashi, Dulhunty and Sakoda (2005) Hum. Mol. Genet. 14, 2189-2200]. In the present study, we explore the ASI region using synthetic peptides corresponding to rabbit RyR1 residues Thr3471-Gly3500 either containing [PASI(+)] or lacking [PASI(-)] the ASI residues. Both peptides increased [3H]ryanodine binding to rabbit RyR1s, increased Ca2+ release from sarcoplasmic reti-culum vesicles and increased single RyR1 channel activity. The peptide PASI(-) was more active in each case than PASI(+). [3H]Ryanodine binding to recombinant ASI(+)RyR1 or ASI(-)-RyR1 was enhanced more by PASI(-) than PASI(+), with the greatest increase seen when PASI(-) was added to ASI(-)RyR1. The activation of the RyR channels is consistent with the hypo-thesis that the peptides interrupt an inhibitory inter-domain inter-action and that PASI(-) is more effective at interrupting this interaction than PASI(+). We therefore suggest that the ASI(-) sequence interacts more tightly than the ASI(+) sequence with its binding partner, so that the ASI(-)RyR1 is more strongly inhibited (less active) than the ASI(+)RyR1. Thus the affinity of the binding partners in this inter-domain interaction may deter-mine the activities of the mature and juvenile isoforms of RyR1 and the stronger inhibition in the juvenile isoform may contribute to the myopathy in DM.

    AB - The aim of the present study was to examine residues that are variably spliced in the juvenile and adult isoforms of the skeletal-muscle RyR1 (type 1 ryanodine receptor). The juvenile ASI(-) splice variant is less active than the adult ASI(+) variant and is overexpressed in patients with DM (myotonic dystrophy) [Kimura, Nakamori, Lueck, Pouliquin, Aoike, Fujimura, Dirksen, Takahashi, Dulhunty and Sakoda (2005) Hum. Mol. Genet. 14, 2189-2200]. In the present study, we explore the ASI region using synthetic peptides corresponding to rabbit RyR1 residues Thr3471-Gly3500 either containing [PASI(+)] or lacking [PASI(-)] the ASI residues. Both peptides increased [3H]ryanodine binding to rabbit RyR1s, increased Ca2+ release from sarcoplasmic reti-culum vesicles and increased single RyR1 channel activity. The peptide PASI(-) was more active in each case than PASI(+). [3H]Ryanodine binding to recombinant ASI(+)RyR1 or ASI(-)-RyR1 was enhanced more by PASI(-) than PASI(+), with the greatest increase seen when PASI(-) was added to ASI(-)RyR1. The activation of the RyR channels is consistent with the hypo-thesis that the peptides interrupt an inhibitory inter-domain inter-action and that PASI(-) is more effective at interrupting this interaction than PASI(+). We therefore suggest that the ASI(-) sequence interacts more tightly than the ASI(+) sequence with its binding partner, so that the ASI(-)RyR1 is more strongly inhibited (less active) than the ASI(+)RyR1. Thus the affinity of the binding partners in this inter-domain interaction may deter-mine the activities of the mature and juvenile isoforms of RyR1 and the stronger inhibition in the juvenile isoform may contribute to the myopathy in DM.

    KW - Alternative Splicing

    KW - Amino Acid Sequence

    KW - Animals

    KW - Exons

    KW - Genetic Variation

    KW - Kinetics

    KW - Molecular Sequence Data

    KW - Muscle, Skeletal

    KW - Rabbits

    KW - Recombinant Proteins

    KW - Ryanodine Receptor Calcium Release Channel

    KW - Sequence Alignment

    KW - Sequence Homology, Amino Acid

    KW - Transfection

    U2 - 10.1042/BJ20060686

    DO - 10.1042/BJ20060686

    M3 - Article

    VL - 401

    SP - 317

    EP - 324

    JO - Biochemical Journal

    JF - Biochemical Journal

    SN - 0264-6021

    IS - 1

    ER -