title: Molecular characterization of Pax6(2Neu) through Pax6(10Neu): an extension of the Pax6 allelic series and the identification of two possible hypomorph alleles in the mouse Mus musculus.
abstract: Phenotype-based mutagenesis experiments will increase the mouse mutant resource, generating mutations at previously unmarked loci as well as extending the allelic series at known loci. Mapping, molecular characterization, and phenotypic analysis of nine independent Pax6 mutations of the mouse recovered in mutagenesis experiments is presented. Seven mutations result in premature termination of translation and all express phenotypes characteristic of null alleles, suggesting that Pax6 function requires all domains to be intact. Of major interest is the identification of two possible hypomorph mutations: Heterozygotes express less severe phenotypes and homozygotes develop rudimentary eyes and nasal processes and survive up to 36 hr after birth. Pax6(4Neu) results in an amino acid substitution within the third helix of the homeodomain. Three-dimensional modeling indicates that the amino acid substitution interrupts the homeodomain recognition alpha-helix, which is critical for DNA binding. Whereas cooperative dimer binding of the mutant homeodomain to a paired-class DNA target sequence was eliminated, weak monomer binding was observed. Thus, a residual function of the mutated homeodomain may explain the hypomorphic nature of the Pax6(4Neu) allele. Pax6(7Neu) is a base pair substitution in the Kozak sequence and results in a reduced level of Pax6 translation product. The Pax6(4Neu) and Pax6(7Neu) alleles may be very useful for gene-dosage studies.
authors.affiliation: Institute of Mammalian Genetics, GSF-Research Center for Environment and Health, Neuherberg D-85764, Germany. favor@gsf.de
authors.forename: J
authors.initials: J
authors.lastname: Favor
authors.forename: H
authors.initials: H
authors.lastname: Peters
authors.forename: T
authors.initials: T
authors.lastname: Hermann
authors.forename: W
authors.initials: W
authors.lastname: Schmahl
authors.forename: B
authors.initials: B
authors.lastname: Chatterjee
authors.forename: A
authors.initials: A
authors.lastname: Neuhäuser-Klaus
authors.forename: R
authors.initials: R
authors.lastname: Sandulache
mesh_terms: D000483:Alleles; D000818:Animals; D015153:Blotting, Western; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D004247:DNA / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D019992:Databases as Topic; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D005123:Eye / Q000196:embryology; D005136:Eye Proteins; D018628:Gene Dosage; D006579:Heterozygote; D018398:Homeodomain Proteins / Q000235:genetics*; D006720:Homozygote; D007438:Introns; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009666:Nose / Q000196:embryology; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; D010641:Phenotype; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors
chemical_list: D018076:DNA, Complementary; D005136:Eye Proteins; D018398:Homeodomain Proteins; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012097:Repressor Proteins; D004247:DNA