Figure 2.

Molecular and biochemical characterization of m4 mutants. (A) Amino acid alignment of a region of the Aldolase protein from various species showing complete conservation in plants, nematodes, insects, zebrafish, and mammals. A single nucleotide change in m4 results in a substitution of Histidine for Arginine in this highly conserved region. (B) Western blot of extracts from heads of flies of the indicated genotypes probed with antibodies against Aldolase or triosephosphate isomerase (Tpi). Steady-state levels of Aldolase are decreased in head extracts of m4 homozygotes and m4/Df hemizygotes. The relative intensity of the Aldolase band in each genotype is shown below the blot. For quantification, Aldolase levels are normalized against the Tpi band for each genotype and these values are shown relative to the levels observed in the control strain (w¹¹¹⁸). (C) Steady-state levels of ATP were measured in extracts of flies of the indicated genotypes as described in the Materials and Methods. ATP levels were normalized to total protein and these values are shown relative to the levels measured in the control strain. Data are presented as the mean of the normalized values ± standard error of the mean. ^**P ≤ 0.01, ^***P ≤ 0.001.