Isolation and functional comparison of Dmyd, the Drosophila homologue of the vertebrate myogenic determination genes, with CMD1.
We have isolated a cDNA clone, called Dmyd for Drosophila myogenic determination gene, from a 0-16 hour Drosophila embryo library that encodes a protein with structural and functional characteristics similar to the members of the vertebrate MyoD family (Paterson et al 1991). Dmyd encodes a polypeptide of 332 amino acids with 82% identity to MyoD in the 41 amino acids of the putative helix-loop-helix region and 100% identity in the 13 amino acids of the basic domain proposed to contain the essential recognition code for muscle specific gene activation. The gene is unique and maps to 95A/B on the right arm of the third chromosome. Low stringency hybridizations indicate Dmyd is not a member of a multigene family, similar to MyoD in vertebrates. Dmyd is a nuclear protein in Drosophila, consistent with its role as a nuclear gene regulatory factor, and is proposed to be a transiently expressed marker for a unique subset of muscle founder cells. We have used an 8kb promoter fragment from the gene, which contains the first 55 amino acids of the Dmyd protein, joined to lac Z to follow myogenic precursor cells into muscle fibers using antibodies to beta-galactosidase and Dmyd. Unlike the myogenic factors in vertebrate muscle cells, Dmyd appears to be expressed at a much lower level in differentiated Drosophila muscles so it cannot be followed continuously as a muscle marker. This is reflected in the loss of expression of Dmyd RNA in 12-24 hour embryos, a major period of early myogenesis, as well as in the undetectable level of the nuclear antigen in primary cultures of embryonic and adult Drosophila muscle. Functional differences between Dmyd and CMD1 are described and explained in terms of a model which may give insight to the nature of homo and heterodimer formation in the bHLH family of proteins.
Paterson
B M
BM
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Shirakata
M
M
Nakamura
S
S
Dechesne
C
C
Walldorf
U
U
Eldridge
J
J
Dübendorfer
A
A
Frasch
M
M
Gehring
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins; D015870:Gene Expression / Q000502:physiology; D017344:Genes, Insect / Q000502:physiology*; D007301:Insect Hormones / Q000235:genetics*; D008969:Molecular Sequence Data; D009124:Muscle Proteins; D009132:Muscles / Q000196:embryology*; D017570:MyoD Protein / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics*; D012313:RNA / Q000032:analysis; D013045:Species Specificity; D014157:Transcription Factors
D029721:Drosophila Proteins; D007301:Insect Hormones; D009124:Muscle Proteins; D017570:MyoD Protein; D009687:Nuclear Proteins; D014157:Transcription Factors; C066817:nau protein, Drosophila; D012313:RNA