Arhgap35^tm1Jset

1.5 kb of sequence, including the start codon and encoding the GTPase domain, was replaced by a PGK-neo cassette. While normal protein was undetected by Western blot analysis of homozygous mutant embryo brain lysates, an aberrant protein generated from an internal start codon was identified at a level similar to that of the wild-type protein. Though sequence encoding the RasGAP binding and RhoGAP domains was retained by the mutant allele, the mutant protein failed to undergo tyrosine phosphorylation or RasGAP association in the brain. Analysis of heterozygous mice indicated improper localization of the mutant protein. Whereas 34% of the wild-type protein was isolated in a detergent-insoluble fraction of brain lysate, only 7% of the mutant protein was present in the same fraction. In addition, study of MEFs indicated differential subcellular localization. (J:65296)