The mouse wellhaarig (we) mutations result from defects in epidermal-type transglutaminase 3 (Tgm3)

• Genetic mapping identifies a small number of candidates for the mouse we4J mutation.
• Sequence analysis of four wellhaarig alleles reveals three distinct Tgm3 mutations.
• Complementation testing shows that two wellhaarig mutations are alleles of Tgm3.
• Analysis of skin mRNA indicates that the we4J transcript is unstable.
• Analysis of skin mRNA indicates that the Tgm3Btlr transcript is aberrantly spliced.

The recessive wellhaarig (we) mutations, named for the wavy coat and curly whiskers they generate in homozygotes, have previously been mapped on mouse Chromosome 2. To further limit the possible location of the we locus, we crossed hybrid (C57BL/6 × AKR)F1, we4J/+ females with AKR, we4J/we4J mutant males to create a large backcross family that was typed for various microsatellite markers and single-nucleotide polymorphisms (SNPs) that distinguish strains AKR and B6. This analysis restricted the location of we4J between sites that flank only one gene known to be expressed in skin: epidermal-type transglutaminase 3 (Tgm3). To test Tgm3 as a candidate for the basis of the wellhaarig phenotype we took two approaches. First, we sequenced all Tgm3 coding regions in mice homozygous for four independent, naturally-occurring wellhaarig alleles (we, weBkr, we3J and we4J) and found distinct defects in three of these mutants. Second, we crossed mice homozygous for an induced mutant allele of Tgm3 (Tgm3Btlr) with mice heterozygous for one of the wellhaarig alleles we possess (we4J or weBkr) to test for complementation. Because the progeny inheriting both a recessive we allele and a recessive Tgm3Btlr allele displayed wavy hair, we conclude that the classic wellhaarig mutations result from defects in Tgm3.