Overexpression of MyoD-inducible lysosomal sialidase (neu1) inhibits myogenesis in C2C12 cells.
Lysosomal sialidase, encoded by neu1, is required for the removal of terminal sialic acid residues from a variety of sialoglycoconjugates. In humans, deficiency of this enzyme results in the inborn error of metabolism sialidosis, characterized by the accumulation of sialoglycoconjugates within the nervous system and in peripheral organs. A subset of sialidosis patients present with symptoms of profound muscle dysfunction, including progressive muscular atrophy. We have previously shown that the 5' regulatory region of murine neu1 is typical of skeletal muscle-specific genes due to the presence of several E-boxes and its responsiveness to stimulation by muscle regulatory factors (MRFs) such as MyoD. Here, we report that sialidase activity is increased 6-fold during the first 24 h of differentiation of C2C12 myoblasts followed by an attenuation to pre-differentiation levels by 48 h. We demonstrate that the lysosomal sialidase promoter is highly upregulated by MyoD through a mechanism that is dependent on the MyoD chromatin remodeling domain. We also show that the sialidase promoter is repressed by activated MEK. Inappropriate overexpression of sialidase 48 h after the onset of differentiation results in downregulation of myogenin as well as myosin heavy chain expression and in a halt of the differentiation cascade. This study indicates that lysosomal sialidase is a potent regulator of the early stages of myogenesis.
Champigny
Marc J
MJ
Department of Biology, McMaster University, 1280 Main St. W. LSB 335, Hamilton, Ontario, Canada.
Perry
Robert
R
Rudnicki
Michael
M
Igdoura
Suleiman A
SA
D000818:Animals; D002454:Cell Differentiation; D002459:Cell Fusion; D002460:Cell Line; D042002:Chromatin Assembly and Disassembly; D015536:Down-Regulation; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005822:Genetic Vectors; D008247:Lysosomes / Q000201:enzymology*; D048369:MAP Kinase Kinase 1 / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D024510:Muscle Development; D017570:MyoD Protein / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology* / Q000201:enzymology; D018008:Myogenin / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D009439:Neuraminidase / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D017570:MyoD Protein; D018008:Myogenin; D048369:MAP Kinase Kinase 1; C482127:Map2k1 protein, mouse; C110128:Neu1 protein, mouse; D009439:Neuraminidase; D018995:Myosin Heavy Chains