title: Altered binding of MYF-5 to FOXE1 promoter in non-syndromic and CHARGE-associated cleft palate.
abstract: BACKGROUND
  Three different homozygous loss-of-function mutations of the Forkhead box E1 (FOXE1) gene have been associated with syndromic cleft palate. Here, we screened the entire promoter region to identify the variations in significant consensus motifs affecting FOXE1 transcription.
  
  
  METHODS
  Genomic DNAs of 35 cleft palate patients, 10 of whom with CHARGE association, 80 unrelated healthy people and 80 unaffected first-degree relatives were analysed by automatic sequencing. The Transcription Element Search System program was employed to identify transcription factor binding sites. The protein-DNA complexes were observed using DNA band-shift assays and oligonucleotide competition analyses. Real-time PCR was used to estimate FOXE1 expression at mRNA level.
  
  
  RESULTS
  In 11 non-syndromic cleft palate patients, a novel non-coding polymorphism (C-->G) in the 5'-untranslated region of FOXE1 was found. The variation fell into a putative consensus sequence for the transcription factor MYF-5 and completely impaired the ability of MYF-5 to bind to its motif, as shown by EMSA experiments. As a consequence, a significantly reduced FOXE1 mRNA expression was observed.
  
  
  CONCLUSIONS
  In 45% of non-syndromic cleft palate patients, a novel homozygous polymorphism that prevented the binding of MYF-5 to FOXE1 promoter and affected the FOXE1 expression was found. As recent data show the role of MYF-5 in the muscle-dependent craniofacial skeletal development and in the fusion of primary palate and secondary palate, the results reported here strongly suggest a more significant involvement of this factor in the cleft palate onset.
authors.affiliation: Department of Neurosurgery, University of Messina, Messina, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Venza
authors.forename: Maria
authors.initials: M
authors.lastname: Visalli
authors.forename: Isabella
authors.initials: I
authors.lastname: Venza
authors.forename: Claudia
authors.initials: C
authors.lastname: Torino
authors.forename: Barbara
authors.initials: B
authors.lastname: Tripodo
authors.forename: Rocco
authors.initials: R
authors.lastname: Melita
authors.forename: Diana
authors.initials: D
authors.lastname: Teti
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D000293:Adolescent; D002648:Child; D002972:Cleft Palate / Q000235:genetics*; D016384:Consensus Sequence / Q000235:genetics; D003596:Cytosine; D004252:DNA Mutational Analysis; D015536:Down-Regulation; D024721:E-Box Elements / Q000235:genetics; D051858:Forkhead Transcription Factors / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics; D006147:Guanine; D006720:Homozygote; D006801:Humans; D009154:Mutation / Q000235:genetics; D051787:Myogenic Regulatory Factor 5 / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D013577:Syndrome; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; C106894:FOXE1 protein, human; D051858:Forkhead Transcription Factors; C497417:MYF5 protein, human; D051787:Myogenic Regulatory Factor 5; D012333:RNA, Messenger; D006147:Guanine; D003596:Cytosine