title: Three new PAX6 mutations including one causing an unusual ophthalmic phenotype associated with neurodevelopmental abnormalities.
abstract: OBJECTIVE
  The PAX6 gene was first described as a candidate for human aniridia. However, PAX6 expression is not restricted to the eye and it appears to be crucial for brain development. We studied PAX6 mutations in a large spectrum of patients who presented with aniridia phenotypes, Peters' anomaly, and anterior segment malformations associated or not with neurological anomalies.
  
  
  METHODS
  Patients and related families were ophthalmologically phenotyped, and in some cases neurologically and endocrinologically examined. We screened the PAX6 gene by direct sequencing in three groups of patients: those affected by aniridia; those with diverse ocular manifestations; and those with Peters' anomaly. Two mutations were investigated by generating crystallographic representations of the amino acid changes.
  
  
  RESULTS
  Three novel heterozygous mutations affecting three unrelated families were identified: the g.572T>C nucleotide change, located in exon 5, and corresponding to the Leucine 46 Proline amino-acid mutation (L46P); the g.655A>G nucleotide change, located in exon 6, and corresponding to the Serine 74 Glycine amino-acid mutation (S74G); and the nucleotide deletion 579delG del, located in exon 6, which induces a frameshift mutation leading to a stop codon (V48fsX53). The L46P mutation was identified in affected patients presenting bilateral microphthalmia, cataracts, and nystagmus. The S74G mutation was found in a large family that had congenital ocular abnormalities, diverse neurological manifestations, and variable cognitive impairments. The 579delG deletion (V48fsX53) caused in the affected members of the same family bilateral aniridia associated with congenital cataract, foveal hypolasia, and nystagmus. We also detected a novel intronic nucleotide change, IVS2+9G>A (very likely a mutation) in an apparently isolated patient affected by a complex ocular phenotype, characterized primarily by a bilateral microphthalmia. Whether this nucleotide change is indeed pathogenic remains to be demonstrated. Two previously known heterozygous mutations of the PAX6 gene sequence were also detected in patients affected by aniridia: a de novo previously known nucleotide change, g.972C>T (Q179X), in exon 8, leading to a stop codon and a heterozygous g.555C>A (C40X) recurrent nonsense mutation in exon 5. No mutations were found in patients with Peters' anomaly.
  
  
  CONCLUSIONS
  We identified three mutations associated with aniridia phenotypes (Q179X, C40X, and V48fsX53). The three other mutations reported here cause non-aniridia ocular phenotypes associated in some cases with neurological anomalies. The IVS2+9G>A nucleotide change was detected in a patient with a microphthalmia phenotype. The L46P mutation was detected in a family with microphthalmia, cataract, and nystagmus. This mutation is located in the DNA-binding paired-domain and the crystallographic representations of this mutation show that this mutation may affect the helix-turn-helix motif, and as a consequence the DNA-binding properties of the resulting mutated protein. Ser74 is located in the PAX6 PD linker region, essential for DNA recognition and DNA binding, and the side chain of the Ser74 contributes to DNA recognition by the linker domain through direct contacts. Crystallographic representations show that the S74G mutation results in no side chain and therefore perturbs the DNA-binding properties of PAX6. This study highlights the severity and diversity of the consequences of PAX6 mutations that appeared to result from the complexity of the PAX6 gene structure, and the numerous possibilities for DNA binding. This study emphasizes the fact that neurodevelopmental abnormalities may be caused by PAX6 mutations. The neuro-developmental abnormalities caused by PAX6 mutations are probably still overlooked in the current clinical examinations performed throughout the world in patients affected by PAX6 mutations.
authors.affiliation: EA n 2502 du Ministère de la Recherche, de l'Enseignement Supérieur et la Technologie, Center de Recherches Thérapeutiques en Ophtalmologie (CERTO), Université René Descartes-Paris V, Faculté de Médecine René Descartes-Site Necker, Paris, France.
authors.forename: Anouk
authors.initials: A
authors.lastname: Dansault
authors.forename: Gabriel
authors.initials: G
authors.lastname: David
authors.forename: Claire
authors.initials: C
authors.lastname: Schwartz
authors.forename: Carolina
authors.initials: C
authors.lastname: Jaliffa
authors.forename: Véronique
authors.initials: V
authors.lastname: Vieira
authors.forename: Guillaume
authors.initials: G
authors.lastname: de la Houssaye
authors.forename: Karine
authors.initials: K
authors.lastname: Bigot
authors.forename: Françise
authors.initials: F
authors.lastname: Catin
authors.forename: Laurent
authors.initials: L
authors.lastname: Tattu
authors.forename: Catherine
authors.initials: C
authors.lastname: Chopin
authors.forename: Philippe
authors.initials: P
authors.lastname: Halimi
authors.forename: Olivier
authors.initials: O
authors.lastname: Roche
authors.forename: Nicole
authors.initials: N
authors.lastname: Van Regemorter
authors.forename: Francis
authors.initials: F
authors.lastname: Munier
authors.forename: Daniel
authors.initials: D
authors.lastname: Schorderet
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Dufier
authors.forename: Cécile
authors.initials: C
authors.lastname: Marsac
authors.forename: Daniel
authors.initials: D
authors.lastname: Ricquier
authors.forename: Maurice
authors.initials: M
authors.lastname: Menasche
authors.forename: Alfred
authors.initials: A
authors.lastname: Penfornis
authors.forename: Marc
authors.initials: M
authors.lastname: Abitbol
mesh_terms: D000015:Abnormalities, Multiple / Q000235:genetics; D000293:Adolescent; D000328:Adult; D000368:Aged; D015783:Aniridia / Q000235:genetics; D000869:Anterior Eye Segment / Q000002:abnormalities; D002386:Cataract / Q000150:complications / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D005124:Eye Abnormalities / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008850:Microphthalmos / Q000150:complications / Q000235:genetics; D008875:Middle Aged; D009154:Mutation; D009421:Nervous System Malformations / Q000235:genetics*; D020417:Nystagmus, Congenital / Q000150:complications / Q000235:genetics; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics*; D010641:Phenotype; D012097:Repressor Proteins / Q000235:genetics*
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; C000605730:PAX6 protein, human; D051761:Paired Box Transcription Factors; D012097:Repressor Proteins