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. 2020 Dec 22;9:e60987. doi: 10.7554/eLife.60987

Figure 1. Allele swap strategy and phenotypic rescue by a diverged version of HOAP.

(A) Using CRISPR/Cas9-mediated transgenesis, we replaced the native coding sequence of cav/HOAP with either a Flag-tagged D. melanogaster coding sequence or instead a Flag-tagged D. yakuba coding sequence. Both coding sequences were intron-less and codon-optimized for D. melanogaster. (B) Mitotic chromosome squashes from larval brains homozygous for HOAP[mel]-Flag or HOAP[yak]-Flag stained with anti-Flag. (C) Mitotic chromosome squashes from larval brains homozygous for HA-HipHop, HOAP[mel]-Flag or HA-HipHop, HOAP[yak]-Flag stained with anti-HA.

Figure 1.

Figure 1—figure supplement 1. Amino acid alignment of HOAP[mel] and HOAP[yak].

Figure 1—figure supplement 1.

The two proteins share less than 75% amino acid identity and even less when indels are taken into account. The only identifiable domain is the N-terminal HMG-box. The dashed box represents the missing sequence from the truncation mutant described in Cenci et al., 2003.
Figure 1—figure supplement 2. Valley of heterozygosity around the cav locus in D. yakuba.

Figure 1—figure supplement 2.

Parameter estimates from Rogers et al., 2015 centered around the cav locus. Theta-pi (A) and Tajima’s D (B) calculated from 20 D. yakuba alleles across 5kb windows sliding 500bp along 200kb (top) and 100kb (bottom).
Figure 1—figure supplement 3. Western Blots of ovaries probed with anti-H3 (control) and anti-Flag to detect Flag-tagged HOAP.

Figure 1—figure supplement 3.

(A) Ovaries hemizygous or homozygous for the transgenes (B) '*' Ovaries from homozygous genotypes used in replicated experimental evolution, '**' genotype not included in downstream experiments.