Malignant-hyperthermia susceptibility is associated with a mutation of the alpha 1-subunit of the human dihydropyridine-sensitive L-type voltage-dependent calcium-channel receptor in skeletal muscle.

Malignant hyperthermia susceptibility (MHS) is characterized by genetic heterogeneity. However, except for the MHS1 locus, which corresponds to the skeletal muscle ryanodine receptor (RYR1) and for which several mutations have been described, no direct molecular evidence for a mutation in another gene has been reported so far. In this study we show that the CACNL1A3 gene encoding the alpha 1-subunit of the human skeletal muscle dihydropyridine-sensitive L-type voltage-dependent calcium channel (VDCC) represents a new MHS locus and is responsible for the disease in a large French family. Linkage analysis performed with an intragenic polymorphic microsatellite marker of the CACLN1A3 gene generated a two-point LOD score of 4.38 at a recombinant fraction of 0. Sequence analysis of the coding region of the CACLN1A3 gene showed the presence of an Arg-His substitution at residue 1086, resulting from the transition of A for G3333, which segregates perfectly with the MHS phenotype in the family. The mutation is localized in a very different part of the alpha 1-subunit of the human skeletal muscle VDCC, compared with previously reported mutations found in patients with hypokalemic periodic paralysis, and these two diseases might be discussed in terms of allelic diseases. This report is the first direct evidence that the skeletal muscle VDCC is involved in MHS, and it suggests a direct interaction between the skeletal muscle VDCC and the ryanodine receptor in the skeletal muscle sarcoplasmic reticulum.

[1]  M. Lehane,et al.  Detection of a novel common mutation in the ryanodine receptor gene in malignant hyperthermia: implications for diagnosis and heterogeneity studies. , 1994, Human molecular genetics.

[2]  F. Couch,et al.  Assignment of the human gene for the alpha 1 subunit of the skeletal muscle DHP-sensitive Ca2+ channel (CACNL1A3) to chromosome 1q31-q32. , 1993, Genomics.

[3]  K. Campbell,et al.  Dystrophin-glycoprotein complex: molecular organization and critical roles in skeletal muscle. , 1995, Current opinion in neurology.

[4]  M. Jeanpierre.  A rapid method for the purification of DNA from blood. , 1987, Nucleic acids research.

[5]  Thomas Deufel,et al.  Exclusion of malignant hyperthermia susceptibility (MHS) from a putative MHS2 locus on chromosome 17q and of the α1, β1, and γ subunits of the dihydropyridine receptor calcium channel as candidates for the molecular defect , 1993 .

[6]  Andrew G. Engel,et al.  Dihydropyridine receptor mutations cause hypokalemic periodic paralysis , 1994, Cell.

[7]  Hanh T. Nguyen,et al.  Enhanced dihydropyridine receptor channel activity in the presence of ryanodine receptor , 1996, Nature.

[8]  M. Leppert,et al.  Sodium channel mutations in paramyotonia congenita and hyperkalemic periodic paralysis , 1993, Annals of neurology.

[9]  P. Powers,et al.  Molecular characterization of the gene encoding the gamma subunit of the human skeletal muscle 1,4-dihydropyridine-sensitive Ca2+ channel (CACNLG), cDNA sequence, gene structure, and chromosomal location. , 1993, The Journal of biological chemistry.

[10]  P. Iaizzo,et al.  Are myotonias and periodic paralyses associated with susceptibility to malignant hyperthermia? , 1990, British journal of anaesthesia.

[11]  B. Britt,et al.  A substitution of cysteine for arginine 614 in the ryanodine receptor is potentially causative of human malignant hyperthermia. , 1991, Genomics.

[12]  M. Phillips,et al.  Ryanodine receptor gene is a candidate for predisposition to malignant hyperthermia , 1990, Nature.

[13]  W. Koch,et al.  Calcium channels from Cyprinus carpio skeletal muscle. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[14]  V. McKusick.  Mendelian inheritance in man , 1971 .

[15]  S. Scherer,et al.  Localization of the gene encoding the alpha 2/delta-subunits of the L-type voltage-dependent calcium channel to chromosome 7q and analysis of the segregation of flanking markers in malignant hyperthermia susceptible families. , 1994, Human molecular genetics.

[16]  C. Doriguzzi,et al.  Mutations in the ryanodine receptor gene in central core disease and malignant hyperthermia , 1993, Nature Genetics.

[17]  J. Weissenbach,et al.  A calcium channel mutation causing hypokalemic periodic paralysis. , 1994, Human molecular genetics.

[18]  B. Bean.  Nitrendipine block of cardiac calcium channels: high-affinity binding to the inactivated state. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[19]  A. Olckers,et al.  Adult muscle sodium channel alpha-subunit is a gene candidate for malignant hyperthermia susceptibility. , 1992, Genomics.

[20]  W. Catterall,et al.  Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[21]  K. Beam,et al.  Restoration of junctional tetrads in dysgenic myotubes by dihydropyridine receptor cDNA. , 1994, Biophysical journal.

[22]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[23]  K. De Jongh,et al.  Characterization of the two size forms of the alpha 1 subunit of skeletal muscle L-type calcium channels. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[24]  F. Hannan,et al.  Cloning and characterization of a calcium channel alpha 1 subunit from Drosophila melanogaster with similarity to the rat brain type D isoform , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[25]  K. De Jongh,et al.  Subunits of purified calcium channels: a 212-kDa form of alpha 1 and partial amino acid sequence of a phosphorylation site of an independent beta subunit. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Denise S Walker,et al.  Direct binding of G-protein βλ complex to voltage-dependent calcium channels , 1997, Nature.

[27]  A. Olckers,et al.  Evidence for the localization of a malignant hyperthermia susceptibility locus (MHS2) to human chromosome 17q. , 1992, Genomics.

[28]  B. Wieringa,et al.  High-resolution physical mapping of four microsatellite repeat markers near the RYR1 locus on chromosome 19q13.1 and apparent exclusion of the MHS locus from this region in two malignant hyperthermia susceptible families. , 1992, Genomics.

[29]  V. Flockerzi,et al.  Primary structure of the receptor for calcium channel blockers from skeletal muscle , 1987, Nature.

[30]  B. Adams,et al.  Regions of the skeletal muscle dihydropyridine receptor critical for excitation–contraction coupling , 1990, Nature.

[31]  M. G. Larach.  Standardization of the Caffeine Halothane Muscle Contracture Test , 1989, Anesthesia and analgesia.

[32]  P. Powers,et al.  The structure of the gene encoding the human skeletal muscle alpha 1 subunit of the dihydropyridine-sensitive L-type calcium channel (CACNL1A3). , 1996, Genomics.

[33]  C. van Broeckhoven,et al.  Mapping of a further malignant hyperthermia susceptibility locus to chromosome 3q13.1. , 1995, American journal of human genetics.

[34]  B. Kay,et al.  A two-motif isoform of the major calcium channel subunit in skeletal muscle , 1992, Neuron.

[35]  A. Stewart,et al.  Genetic mapping of the β1- and γ-subunits of the human skeletal muscle L-type voltage-dependent calcium channel on chromosome 17q and exclusion as candidate genes for malignant hyperthermia susceptibility , 1993 .

[36]  G. Lathrop,et al.  Easy calculations of lod scores and genetic risks on small computers. , 1984, American journal of human genetics.

[37]  M. Farrall,et al.  Localization of the malignant hyperthermia susceptibility locus to human chromosome 19ql2–13.2 , 1990, Nature.

[38]  K. Itagaki,et al.  Identification of 1,4‐dihydropyridine binding domains within the primary structure of the α1 subunit of the skeletal muscle L‐type calcium channel , 1993, FEBS letters.

[39]  W. Catterall,et al.  Structure and function of voltage-sensitive ion channels. , 1988, Science.

[40]  W. Kalow.  Inheritance of Malignant Hyperthermia—A Review of Published Data , 1987 .

[41]  M. Phillips,et al.  A mutation in the human ryanodine receptor gene associated with central core disease , 1993, Nature Genetics.

[42]  M. Pinaud,et al.  Malignant hyperthermia in a patient with hypokalemic periodic paralysis. , 1994, Anesthesia and analgesia.

[43]  N. Monnier,et al.  Mutation screening of the RYR1 gene in malignant hyperthermia: detection of a novel Tyr to Ser mutation in a pedigree with associated central cores. , 1994, Genomics.

[44]  P. Powers,et al.  Assignment of the human gene for the beta subunit of the voltage-dependent calcium channel (CACNLB1) to chromosome 17 using somatic cell hybrids and linkage mapping. , 1993, Genomics.

[45]  S. Scherer,et al.  Localization of the gene encoding the α2/δ subunit (CACNL2A) of the human skeletal muscle voltage-dependent Ca2+ channel to chromosome 7q21- q22 by somatic cell hybrid analysis , 1994 .

[46]  P. Powers,et al.  Cloning of the human skeletal muscle alpha 1 subunit of the dihydropyridine-sensitive L-type calcium channel (CACNL1A3). , 1994, Genomics.

[47]  T. Meitinger,et al.  Evidence for genetic heterogeneity of malignant hyperthermia susceptibility. , 1992, American journal of human genetics.

[48]  T. Snutch,et al.  Crosstalk between G proteins and protein kinase C mediated by the calcium channel α1 subunit , 1997, Nature.

[49]  K. Campbell,et al.  Calcium channel β-subunit binds to a conserved motif in the I–II cytoplasmic linker of the α1-subunit , 1994, Nature.