Prediction and characterisation of a highly conserved, remote and cAMP responsive enhancer that regulates Msx1 gene expression in cardiac neural crest and outflow tract.
Double knockouts of the Msx1 and Msx2 genes in the mouse result in severe cardiac outflow tract malformations similar to those frequently found in newborn infants. Despite the known role of the Msx genes in cardiac formation little is known of the regulatory systems (ligand receptor, signal transduction and protein-DNA interactions) that regulate the tissue-specific expression of the Msx genes in mammals during the formation of the outflow tract. In the present study we have used a combination of multi-species comparative genomics, mouse transgenic analysis and in-situ hybridisation to predict and validate the existence of a remote ultra-conserved enhancer that supports the expression of the Msx1 gene in migrating mouse cardiac neural crest and the outflow tract primordia. Furthermore, culturing of embryonic explants derived from transgenic lines with agonists of the PKC and PKA signal transduction systems demonstrates that this remote enhancer is influenced by PKA but not PKC dependent gene regulatory systems. These studies demonstrate the efficacy of combining comparative genomics and transgenic analyses and provide a platform for the study of the possible roles of Msx gene mis-regulation in the aetiology of congenital heart malformation.
Miller
Kerry Ann
KA
School of Medical Sciences, Foresterhill, University of Aberdeen, Aberdeen AB25 2ZD, UK.
Davidson
Scott
S
Liaros
Angela
A
Barrow
John
J
Lear
Marissa
M
Heine
Danielle
D
Hoppler
Stefan
S
MacKenzie
Alasdair
A
D000818:Animals; D001483:Base Sequence; D019295:Computational Biology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000196:embryology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051957:MSX1 Transcription Factor / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009432:Neural Crest / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction / Q000502:physiology; D013045:Species Specificity
D017931:DNA Primers; D051957:MSX1 Transcription Factor; C484084:Msx1 protein, mouse; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C