Fig. 2. Structure and expression of Bchs protein. (A) The domains of
Drosophila Bchs and human Alfy are similar in organization and
sequence. The percentage amino acid identity between domains is indicated,
including homology for the extended portion of the protein called CRAB
(Conserved Region in Alfy and Bchs; shown in gray and not shown to scale to
conserve space). Mutations in bchs alleles are marked (nonsense,
stars; missense, triangles). (B) In a stage 13 embryo, bchs
mRNA is highly expressed in embryonic CNS, including brain and ventral nerve
cord, and in the salivary glands (arrowhead). A sense control probe showed no
signal above background (data not shown). (C) Analysis of murine mRNA
indicates mouse Alfy is widely expressed, but enriched in adult brain.
(D) Protein blot probed with antisera raised to amino acids 2237-2590
of Bchs (arrow indicates Bchs; just below Bchs, the anti-sera also detects a
crossreacting protein, whose size and intensity are unaffected in multiple
bchs truncation alleles). GMR-GAL4;EP-bchs animals express
more Bchs protein than controls, including EP-bchs. Overexpression is
abolished by the nonsense mutation in bchs17
(GMR-GAL4;bchs17/+). bchs12,
bchs17 and bchs58 express no detectable
Bchs, while bchs8 still produces substantial amounts of
protein. Antibody against Elav demonstrates equal loading. VNC, ventral nerve
cord.