(A) Sequences of the first 34 amino acids of wild-type Yki and the
YkiAcidA allele. Asterisks indicate acidic amino acids.
(B–E′) Mutation of acidic residues at the N-terminus of Yki
blocks its ability to activate myosin. S2 cells expressing membrane-associated
wild-type Myr-Yki (B–B‴) and Myr-YkiAcidA
(C–C‴). Myr-YkiAcidA fails to activate Sqh. White
arrows indicate transfected cells. Wing imaginal discs expressing
Myr-YkiAcidA also fail to activate Sqh (D–E′).
The two transgenes were integrated at the same landing site to ensure comparable
expression levels.
(F–K) The myosin-activating function of Yki is required for proper
growth. Homozygous ykiAcidA adults are smaller than
controls (F). Representative images show that adult female wings from animals
homozygous for the ykiAcidA allele are undergrown
(G–H). Larval wing imaginal discs are similarly undergrown
(I–K), with ykiAcidA/ykiB5
(ykiAcidA/−) animals displaying the
strongest phenotype (the latter genotype does not survive to adults).
Quantification of adult wing and larval wing disc sizes of the indicated
genotypes confirms these differences (L–M). Data are represented as mean
± SEM. Asterisks represent statistical significance of the difference
between selected groups (**** p<0.0001,
One-way ANOVA and Tukey’s HSD test, n = number of wings
[L]/wing discs [M]).
(N) Model of feedback regulation of the Hippo pathway. Two negative feedback
mechanisms have been proposed in the regulation of the Hippo pathway (red
circles). One is mediated through transcriptional upregulation of upstream
components, including Mer and Ex, and the other is a consequence of
morphogen-driven growth at the center of tissues and the resulting compression
of those cells. We propose a positive feedback mechanism (green circle) in which
cortical Yki at the AJR promotes myosin activation and increased tension via
Strn-Mlck, which in turn further activates Yki by suppressing the Hippo pathway.
See the Discussion for more details.
See also Figure S7.