THE STRUCTURAL ORGANIZATION OF THE HUMAN SKELETAL-MUSCLE RYANODINE RECEPTOR (RYR1) GENE

The RYR1 gene encoding the Ca2+ release channel of human skeletal musc le sarcoplasmic reticulum has been cloned and exon/intron boundaries h ave been determined, together with a minimum of 30 bp of intron sequen ce flanking each splice junction. The gene contains 106 exons, of whic h two are alternatively spliced. The length of the gene, determined by the alignment of 16 genomic phage clones, a cosmid. clone, and severa l long polymerase chain reaction products, is approximately 160 kb. Ex ons range from 15 to 813 bp, while introns range from 85 to about 16,0 00 bp, Analysis of the gene has confirmed published errors in the huma n RYR1 cDNA and confirmed the structure of two alternatively spliced e xons. The numbering of the nucleotides comprising the RYR1 cDNA and th e numbering of amino acids encoded by them were corrected to account f or these earlier errors and omissions. Analysis of 2.4 kb of the 5' up stream sequence indicated the presence of a CCAAT box and several Spl binding sites between nucleotides -200 and -60 br,, flanking the propo sed transcription start site at -130 bp. Several other potential trans cription factor binding sites were identified throughout the 5' sequen ce. Knowledge of the structure of the RYR1 gene will provide an invalu able resource for the discovery of mutations in the gene that are caus al of human malignant hyperthermia and central core disease. (C) 1990 Academic Press, Inc.