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Fig. 4. EMS induced Chro alleles. (A) Diagram of the wild-type Chromator protein and the potential truncated protein products of the EMS induced Chro alleles, Chro⁷¹ and Chro⁶¹². The Chro⁷¹ allele is comprised of a G to A nucleotide change at nucleotide position 402 of the Chro coding sequence that introduces a premature stop codon resulting in a truncated 71 amino acid protein. The Chro⁶¹² allele consists of a C to T nucleotide change at nucleotide position 2024 that introduces a premature stop codon resulting in a truncated 612 amino acid protein that retains the chromodomain. (B) Chromator protein expression in Chro⁷¹/Chro⁶¹² mutant third-instar larvae compared with wild type larvae. The immunoblots were labeled with the C-terminal Chromator mAb 6H11 and with anti-tubulin antibody as a loading control. Full-length Chromator is detected as a 130 kDa protein by mAb 6H11 in wild-type larvae; however, no full-length Chromator is detectable in the mutant larvae. The relative migration of molecular weight markers is indicated to the left of the immunoblots in kDa. (C,D) Polytene chromosome preparations from third-instar larvae were labeled with Hoechst to visualize the chromatin. Preparations are shown from a wild-type female larvae (C) and from a female Chro⁷¹/Chro⁶¹² mutant larvae (D). Reduced levels of wild-type Chromator protein have a severe effect on the structure and organization of larval polytene chromosomes. Note the disruption and misalignment of interband and banded regions and the extensive coiling and folding of the chromosome arms in Chro⁷¹/Chro⁶¹² mutant chromosomes (D).

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