Expression and DNA sequence analysis of a human embryonic skeletal muscle myosin heavy chain gene.
Vertebrate myosin heavy chains (MHC) are represented by multiple genes that are expressed in a spatially and temporally distinct pattern during development. In order to obtain molecular probes for developmentally regulated human MHC isoforms, we used monoclonal antibodies to screen an expression cDNA library constructed from primary human myotube cultures. A 3.4 kb cDNA was isolated that encodes one of the first MHCs to be transcribed in human skeletal muscle development. A portion of the corresponding gene encoding this isoform has also been isolated. Expression of this embryonic MHC is a hallmark of muscle regeneration after birth and is a characteristic marker of human muscular dystrophies. During normal human development, expression is restricted to the embryonic period of development prior to birth. In primary human muscle cell cultures, devoid of other cell types, mRNA accumulation begins as myotubes form, reaches a peak 2 days later and declines to undetectable levels within 10 days. The expression of the protein encoded by the embryonic skeletal MHC gene follows a similar time course, lagging behind the mRNA by approximately two days. Thus, expression of the human embryonic gene is efficiently induced and then repressed in cultured muscle cells, as it is in muscle tissue. The study of the regulation of a human MHC isoform with a central role in muscle development and in muscle regeneration in disease states is therefore amendable to analysis at a molecular level.
Karsch-Mizrachi
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Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461.
Travis
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M
Blau
H
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Leinwand
L A
LA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D015246:Deoxyribonuclease EcoRI; D005786:Gene Expression Regulation; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009132:Muscles / Q000196:embryology* / Q000378:metabolism; D009218:Myosins / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D015342:DNA Probes; D004247:DNA; D015246:Deoxyribonuclease EcoRI; D009218:Myosins