Characterization of the mouse lysosomal sialidase promoter.
Lysosomal sialidase is required for the catabolism of sialoglycoconjugates such as gangliosides and deficiency in this enzyme results in the autosomal recessive disease sialidosis. Furthermore, we have shown that overexpression of human sialidase is sufficient to clear accumulated ganglioside in Tay-Sachs neuroglia [Hum. Mol. Genet. 8 (1999) 1111]. In this paper, we have characterized the 5' regulatory region of the mouse lysosomal sialidase gene in order to understand the molecular mechanisms regulating its expression. We used bioinformatic approaches to identify a transcriptional initiation site at -45 bp relative to the ATG and significant sequence homology with the rat and human promoters. Expression by the promoter was found to be cell-type restricted and required at least 750 bp upstream of the ATG for high-level expression. DNAse I footprinting analysis and reporter gene assays indicated that the promoter is responsive to Sp-1. We discovered a CCAAT box and four E-boxes within the mouse upstream region and demonstrated that CCAAT displacement protein as well as the muscle regulatory factors MyoD and Myf-5 influence sialidase expression. Taken together, these results identify cis- and trans-acting factors involved in the regulation of sialidase and point to mechanisms of gene upregulation.
Champigny
Marc J
MJ
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Johnson
Matthew
M
Igdoura
Suleiman A
SA
D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D017124:Conserved Sequence / Q000235:genetics; D018983:DNA Footprinting; D004268:DNA-Binding Proteins; D024202:Electrophoretic Mobility Shift Assay; D019143:Evolution, Molecular; D051537:Hepatocyte Nuclear Factor 1; D051538:Hepatocyte Nuclear Factor 1-alpha; D051539:Hepatocyte Nuclear Factor 1-beta; D018398:Homeodomain Proteins; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D041681:NIH 3T3 Cells; D009439:Neuraminidase / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D022762:CCAAT-Enhancer-Binding Proteins; C080632:CUX1 protein, human; C000624101:Cux1 protein, mouse; D004268:DNA-Binding Proteins; C496487:HNF1A protein, human; C496495:HNF1B protein, human; D051538:Hepatocyte Nuclear Factor 1-alpha; C496488:Hnf1a protein, mouse; C496489:Hnf1a protein, rat; C496496:Hnf1b protein, mouse; D018398:Homeodomain Proteins; D017570:MyoD Protein; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D016329:Sp1 Transcription Factor; D014157:Transcription Factors; D051537:Hepatocyte Nuclear Factor 1; D051539:Hepatocyte Nuclear Factor 1-beta; D008156:Luciferases; D009439:Neuraminidase