DEV01821 Supplementary Material

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• Supplemental Figure 1 (Adobe PDF) -

  Fig. S1. The PAT-related transporter family. (A) Figure shows an alignment of human PAT1-4, CG1139, PATH and PATH-like sequences from the mosquito Anopheles gambiae (XM_308237) and the Western honeybee Apis mellifera (XP_396451). The line-up is annotated according to the 11 transmembrane domain model (Sagne et al., 2001; Boll et al., 2004) and reveals highest levels of homology in some of these domains (particularly 1, 4, 6 and 10). There are at least eight other PAT-related genes in flies. (B,C) Homology tables derived form BLAST searches showing % amino acid identity and similarity (parentheses) between different human and Drosophila PAT-related transporters (B) and PATH-like transporters (C). Human PATs share between 68% (PAT1 versus PAT2) and 46% (PAT3 versus PAT4) identity. PATH, XM_308237 and XP_396451 also share high levels of identity. Again, the greatest regions of homology are in the putative transmembrane domains, and include the region marked TM1 in A. Although we do not believe that PATH-like molecules insert with a different topology in the plasma membrane, TM1 is interestingly not predicted to be inserted into the cell membrane for these molecules.

  References

  Boll, M., Daniel, H. and Gasnier, B. (2004). The SLC36 family: proton-coupled transporters for the absorption of selected amino acids from extracellular and intracellular proteolysis. Pflugers Arch. 447, 776-779.

• Supplemental Figure 2 -

  Fig. S2. FOXO activity is specifically modulated by upstream components of the InR pathway. GMR-GAL4 was used to drive expression of several transgenes in the differentiating cells of the eye in the presence (B,D,F,H,J,L,N) or absence (C,E,G,I,K,M) of foxo^GS9928. Overexpression of foxo^GS9928 alone produced a marked reduction in eye size, particularly in the posteroentral region (arrow in B), relative to control eyes (A). The foxo-induced phenotype in B was strongly enhanced by a dominant-negative form of PI3-kinase, UAS-Dp110^D954A (D), which by itself produces a mild reduction in ommatidial growth (C). Conversely, the foxo phenotype was strongly suppressed by co-overexpressing Dp110 (F) or Akt1 (H), which increase InR signalling and therefore induce overgrowth in the absence of foxo (E and G, respectively). By contrast, positive regulators of growth that are not thought to modulate InR signalling, including ban^EP3622 (I) (Brennecke et al., 2003), UAS-CycD UAS-cdk4 (K) (Datar et al., 2000) and a dominant negative form of hippo, UAS-hpo^K71R (M) (Wu et al., 2003), fail to fully suppress either the ventral reduction phenotype or the general growth inhibition associated with foxo^GS9928 overexpression (J,L,N, respectively). All panels show adult males, as they produced stronger foxo^GS9928 genetic interaction phenotypes. The GMR-GAL4-induced foxo phenotype is particularly sensitive to reduced nutrient conditions, so flies were reared in uncrowded conditions.

  References

  Brennecke, J., Hipfner, D. R., Stark, A., Russell, R. B. and Cohen, S. M. (2003). bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113, 25-36.

  Datar, S. A., Jacobs, H. W., de la Cruz, A. F., Lehner, C. F. and Edgar B. A. (2000). The Drosophila cyclin D-Cdk4 complex promotes cellular growth. EMBO J. 19, 4543-4554.

  Wu, S., Huang, J., Dong, J. and Pan, D. (2003). hippo encodes a Ste-20 family protein kinase that restricts cell proliferation and promotes apoptosis in conjunction with salvador and warts. Cell 114, 445-456.

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