Isolation and nucleotide sequence of human liver glycogen debranching enzyme mRNA: identification of multiple tissue-specific isoforms

• Published in: Gene 1997-09, Vol.197 (1), p.389-398
• Main Authors: Bao, Yong, Yang, Bing-Zhi, Dawson Jr, Thomas L., Chen, Yuan-Tsong
• Format: Article
• Language: English
• Subjects: Alternative splicing; Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Complementary - genetics; Gene expression; Glycogen debranching enzyme; Glycogen Debranching Enzyme System - genetics; Glycogen storage disease; Humans; Liver - chemistry; Liver - enzymology; Molecular Sequence Data; Muscles - chemistry; Organ Specificity; RNA, Messenger - analysis; RNA, Messenger - genetics; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(97)00291-6

Glycogen storage disease type III (GSD-III) is caused by a deficiency of glycogen debranching enzyme (AGL) activity. Patients are found to have deficient AGL activity in both muscle and liver, and also enzyme deficiency in the liver, but not in muscle. To determine the molecular basis of enzymatic variability in GSD-III and to elucidate the mechanism for control of tissue-specific expression of AGL, we previously cloned and sequenced the human muscle AGL cDNA. Here we report the isolation and nucleotide sequence of liver AGL cDNA and the tissue distribution of the isoform mRNAs. The predominant form of human liver AGL cDNA (isoform 1) contained 400 bp of 5′ untranslated region, 4596 bp of coding region, and 2371 bp of 3′ untranslated region. The liver AGL mRNA sequence was identical to the previously published muscle sequence (isoform 5) for most of the length, except for the 5′ end, in which the liver sequence diverged completely from the muscle sequence. The divergence began with the transcription start point and extended 82 nucleotides downstream from the translation initiation codon. Six isoforms of AGL mRNA were identified and sequenced from liver and muscle. These isoforms differed only at the 5′ end. Tissue distribution studies showed that liver, kidney and lymphoblastoid cells expressed predominantly isoform 1; whereas muscle and heart expressed not only isoform 1, but also muscle-specific isoform mRNAs (isoforms 2, 3 and 4). Defining tissue-specific AGL isoform mRNAs is an important step toward understanding the molecular basis of enzymatic variability in GSD-III.