Summary of Drosophila kae1 (CG4933) mutant alleles. (A) An alignment of fruit fly, human, yeast, and archaeal orthologs of Kae1. Functionally/structurally defined regions are highlighted, including the binding regions for Pcc1 and Bud32, and the ATP- and ion-binding regions. All Black Spot alleles recovered from our screen revealed that point mutations affected well-conserved residues. Notably, none of the mutations directly affects a known functional site, although a few mutations are in the vicinity of such regions. The noncomplementing allele l(3)72Fb[331], identified from a public stock collection, bears a nonsense mutation in kae1. Note that 1A2 and 1A19 induce the same coding change, even though these were isolated from different chromosomes in the screen. (B) qRT-PCR analysis of kae1 from various homozygous or hemizygous mutants. All of our point alleles maintain substantial levels of kae1 transcripts, although several of them are reduced by 20%–30%. The pBac insertion into the 5′ UTR of kae1, f[10978], appears to be a null allele as no transcript was detected. (C) Agarose gel analysis of qRT-PCR amplicons confirms the absence of kae1 transcripts in f[10978]/Df larvae.
