A novel protein found in the I bands of myofibrils is produced by alternative splicing of the DLST gene
| Autor: | Shigeo Ohta, Akinori Kimura, Kyoko Nakano, Takuro Arimura, Sadayuki Matuda, Hiroaki Takekura |
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| Rok vydání: | 2010 |
| Předmět: |
Sarcomeres
DNA Complementary Blotting Western Molecular Sequence Data Biophysics Biochemistry Gene Expression Regulation Enzymologic Mice Exon Myofibrils Complementary DNA Animals Humans Amino Acid Sequence RNA Messenger Muscle Skeletal Molecular Biology Peptide sequence Gene Base Sequence Reverse Transcriptase Polymerase Chain Reaction Chemistry Alternative splicing Sequence Analysis DNA Immunohistochemistry Molecular biology Stop codon Rats Isoenzymes Mice Inbred C57BL Alternative Splicing Open reading frame RNA splicing Electrophoresis Polyacrylamide Gel Acyltransferases |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - General Subjects. 1800:31-39 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/j.bbagen.2009.10.003 |
| Popis: | Background It is not known if the dihydrolipoamide succinyltransferase ( DLST ) gene, a mitochondrial protein, undergoes alternative splicing. We identified an uncharacterized protein reacting with an anti-DLST antibody in the I bands of myofibrils in rat skeletal muscle. Methods Immunocytochemical staining with an anti-DLST antibody, the purification and amino acid sequence analysis of the protein, and the isolation and sequencing of the protein's cDNA were carried out to clarify the properties of the protein and its relationship to the DLST gene. Results A pyrophosphate concentration >10 mM was necessary to extract the protein from myofibrils in the presence of salt with a higher concentration than 0.6 M, at an alkaline pH of 7.5–8.0. The protein corresponded to the amino acid sequence of the C-terminal side of DLST. The cDNAs for this protein were splicing variants of the DLST gene, with deletions of both exons 2 and 3, or only exon 2 or 3. These variants possessed an open reading frame from an initiation codon in exon 8 of the DLST gene to a termination codon in exon 15, generating a protein with a molecular weight of 30 kDa. Conclusions The DLST gene undergoes alternative splicing, generating the protein isolated from the I bands of myofibrils. General significance The DLST gene produces two different proteins with quite different functions via alternative splicing. |
| Databáze: | OpenAIRE |
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