| 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. Pax64Neu 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 α-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 Pax64Neu allele. Pax67Neu is a base pair substitution in the Kozak sequence and results in a reduced level of Pax6 translation product. The Pax64Neu and Pax67Neu alleles may be very useful for gene-dosage studies. |
| Keywords: |
gene mutation; mutation; exons; nonhuman; protein domain; mouse; phenotype; animals; mice; mus; amino acid substitution; embryo; gene locus; alleles; introns; eye proteins; homeodomain proteins; heterozygote; mice, inbred c57bl; time factors; mus musculus; blotting, western; dna; reverse transcriptase polymerase chain reaction; nucleotide sequence; homozygote; rna translation; protein biosynthesis; dna sequence; models, molecular; mutagenesis, site-directed; protein structure, tertiary; transcription factor pax6; homeodomain protein; gene dosage; dna binding; repressor proteins; mutagenesis; monomer; eye; chromosome mapping; electrophoresis, polyacrylamide gel; paired box transcription factors; dna, complementary; oncogene neu; mice, inbred c3h; crosses, genetic; alpha helix; nose; databases; allelism; priority journal; article; musculus
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