Altered expression of apicobasal polarity factors is associated with cancer in vertebrates and tissue overgrowth in model invertebrates, yet mechanisms by which these factors affect growth regulatory pathways are not well defined. We have previously identified the Drosophila melanogaster gene tsg101 (tumor susceptibility gene-101) as a factor that is required to simultaneously maintain tissue polarity and suppress tumor-like overgrowth of the larval imaginal discs, which are simple epithelial organs that give rise to most adult structures. We have shown that loss of tsg101 blocks endolysosomal turnover of the key apical membrane determinant Crumbs, and that this excess Crumbs protein accumulates at the apical membrane, spreads ectopically onto the basolateral membrane, and accumulates in late endosomes. As Crumbs nucleates an apical membrane- associated complex that interacts physically and functionally with the Par/aPKC and Scribble/Dlg polarity complexes, these defects in Crumbs regulation provide a ready explanation of the effect of tsg101 loss on epithelial polarity and architecture. Notably, over expressed Crumbs also drives substantial imaginal disc overgrowth, suggesting that this polarity factor has a second, more direct oncogenic role in tsg101 cells. In published work, we have found that the cytoplasmic tail of Crumbs contains a previously unrecognized growth-regulatory motif through which it interacts with and regulates the expanded protein, which is a key component of the conserved Hippo/Mst2 tumor suppressor pathway. These data show that Crumbs is a multi-functional protein capable of integrating junctional polarity signals with the well-established Hippo/Mst2 growth-regulatory pathway, and those lesions that disrupt Crumbs trafficking and localization may have very direct effects on the activity of this pathway. Our objectives in this proposal are to test the role of this newly discovered Crumbs-Hpo/Mst2 link in the excessive growth of tsg101 mutant tumors, and to identify novel components of the Crumbs-Hpo/Mst2 pathway from among a small collection of mutations we have identified as dominant-modifiers of Crumbs driven disc overgrowth. In particular, we will focus our analysis on the taiman gene, which encodes the fly homolog of the human Amplified in Breast Cancer-1 (AIB-1) oncogene, and that we believe acts as a transcriptional coactivator for the Crumbs-Hpo/Mst2 pathway. We will use standard genetic and molecular techniques in Drosophila organs and cultured cells to carry out the proposed studies.

Public Health Relevance

Loss of the human gene Tumor Susceptibility Gene-101 (TSG101) has been suggested to promote formation of cancer, although mechanisms by which this might occur are not well understood. We have found that the fruit fly version of the TSG101 controls a protein called Crumbs, and that Crumbs in turn directly regulates the activity of a very well established cancer-regulatory pathway called the Hippo/Mst2 pathway. We propose to study this Crumbs-Hpo/Mst2 pathway in flies, and to determine how it contributes to the 'cancerous'growth of fly tumors and, by extension, human tumors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA123368-08
Application #
8467684
Study Section
Intercellular Interactions (ICI)
Program Officer
Watson, Joanna M
Project Start
2006-08-01
Project End
2016-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
8
Fiscal Year
2013
Total Cost
$206,920
Indirect Cost
$73,423
Name
Emory University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Barron, Daniel A; Moberg, Kenneth (2016) Inverse regulation of two classic Hippo pathway target genes in Drosophila by the dimerization hub protein Ctp. Sci Rep 6:22726
Zhang, Can; Robinson, Brian S; Xu, Wenjian et al. (2015) The ecdysone receptor coactivator Taiman links Yorkie to transcriptional control of germline stem cell factors in somatic tissue. Dev Cell 34:168-80
Kelly, Seth M; Leung, Sara W; Pak, ChangHui et al. (2014) A conserved role for the zinc finger polyadenosine RNA binding protein, ZC3H14, in control of poly(A) tail length. RNA 20:681-8
Kagey, Jacob D; Brown, Jordan A; Moberg, Kenneth H (2012) Regulation of Yorkie activity in Drosophila imaginal discs by the Hedgehog receptor gene patched. Mech Dev 129:339-49
Robinson, Brian S; Moberg, Kenneth H (2011) Drosophila endocytic neoplastic tumor suppressor genes regulate Sav/Wts/Hpo signaling and the c-Jun N-terminal kinase pathway. Cell Cycle 10:4110-8
Gilbert, M Melissa; Tipping, Marla; Veraksa, Alexey et al. (2011) A screen for conditional growth suppressor genes identifies the Drosophila homolog of HD-PTP as a regulator of the oncoprotein Yorkie. Dev Cell 20:700-12
Beam, Carolyn K; Moberg, Kenneth (2010) The gang of four gene regulates growth and patterning of the developing Drosophila eye. Fly (Austin) 4:104-16
Robinson, Brian S; Huang, Juang; Hong, Yang et al. (2010) Crumbs regulates Salvador/Warts/Hippo signaling in Drosophila via the FERM-domain protein Expanded. Curr Biol 20:582-90
Gilbert, M Melissa; Robinson, Brian S; Moberg, Kenneth H (2009) Functional interactions between the erupted/tsg101 growth suppressor gene and the DaPKC and rbf1 genes in Drosophila imaginal disc tumors. PLoS One 4:e7039
Gilbert, M Melissa; Beam, Carolyn K; Robinson, Brian S et al. (2009) Genetic interactions between the Drosophila tumor suppressor gene ept and the stat92E transcription factor. PLoS One 4:e7083