title: The Role of hsa-miR-548l Dysregulation as a Putative Modifier Factor for Glaucoma-Associated FOXC1 Mutations.
abstract: Mutations of the FOXC1 transcription factor are involved in a variety of autosomal dominant ocular anterior segment defects, ranging from Axenfeld-Rieger malformations to isolated glaucoma in some patients. In this study we have evaluated the possible role of the c.*734A>T FOXC1 variant as a modifier factor of the activity of two FOXC1 mutations previously identified in families primarily affected by dominant glaucoma (haplotypes p.G447_G448insDG-c.*734A>T and p.I126S-c.*734A>T). Previous bioinformatic analyses indicated that the c.*734A>T variant is located in a potential target sequence for hsa-miR-548l. Co-expression of this miRNA with a reporter cDNA construct in which the wild-type 3'UTR sequence of FOXC1 was fused to the 3'-end of the firefly luciferase coding region, led to approximately 20% decreased luciferase activity compared to the controls, confirming the presence of a target sequence for hsa-miR-548l. In contrast, this miRNA did not show any effect on the luciferase activity associated with the mutant 3'UTR FOXC1 sequence, showing that it resulted in a loss-of-function of the has-miR-548l target sequence. In addition, functional evaluation of the two glaucoma-associated haplotypes revealed increased protein levels and transactivation, compared to the corresponding individual coding mutations (approximately 1.2-fold on average). These data support the role of hsa-miR-548l as a regulator of FOXC1 translation and provide evidence for the c.*734A>T variant as a modifier factor for the activity of coding glaucoma-associated FOXC1 mutations.
authors: Cristina
authors: C
authors: Medina-Trillo
authors: José-Daniel
authors: JD
authors: Aroca-Aguilar
authors: Jesús-José
authors: JJ
authors: Ferre-Fernández
authors: Carmen-Dora
authors: CD
authors: Méndez-Hernández
authors: Laura
authors: L
authors: Morales
authors: Julián
authors: J
authors: García-Feijoo
authors: Department of Genetics, Faculty of Medicine, Castilla-La Mancha University, Albacete, Spain. julio.escribano@uclm.es.
authors: Julio
authors: J
authors: Escribano
mesh_terms: D001483:Base Sequence; D005260:Female; D051858:Forkhead Transcription Factors / Q000235:genetics*; D005786:Gene Expression Regulation; D005901:Glaucoma / Q000235:genetics*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics
chemical_list: C112743:FOXC1 protein, human; D051858:Forkhead Transcription Factors; C561887:MIRN548 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger