In this amended PPG Application, 6 NIH-funded Pis propose interactive research focusing on the trafficking of membrane transport proteins in polarized epithelial cells from the small intestine, liver and kidney. Three Projects and 3 supporting Cores are proposed. The proteins, each of which contribute to human diseases, are the BB Na/H exchanger NHE3 (Project 1, PI: M. Donowitz), the intracellular Cl channel CIC-5 (Project 2, PI: S. Guggino, co-investigator W. Guggino), and the Cu-ATPases 7A (Menkes protein) and 7B (Wilson's protein) (Project 3, PI: A. Hubbard). The common questions being addressed are: What are the mechanisms by which these proteins are trafficked? What are the roles of these proteins in the trafficking of other polarized membrane proteins in the same cells? What are the nature of the intracellular compartments in which these proteins distribute? The Cores are an Administrative Core (Core A), a Cell Culture/Adeno and Lentivirus Preparation Core (Core B, Director M. Tse), and a Two-Photon Microscopy and Confocal FRET/FRAP Imaging Core (Core C, Director O. Kovbasnjuk). Project 1, Knockout Models of PDZ Proteins and NHE3 Regulation, PI: M. Donowitz. The hypothesis will be tested that multiple BB PDZ proteins form multiprotein NHE3 complexes which are involved dynamically in NHE3 regulation. BB PDZ domain protein knockout mice and knockdowns (shRNAi) in Caco-2 cells will examine the role of these PDZ proteins in NHE3 regulation, complex size and associated proteins. Project 2, CIC-5 in Endocytosis and NHE3/NaPi-lla Trafficking, PI: S. Guggino. A knockout mouse model of CIC-5 which mimics Dent's disease will be studied for effects on BB endocytosis and exocytosis, trafficking of megalin, NHE3, and NaPi-lla. Whether CIC-3 and/or -4 can substitute for CIC-5 and whether acidification of endosomal pH reconstitutes trafficking effects of a CIC-5 mutant which lacks Cl channel activity but has CI/H exchange activity will test which CIC-5 function is needed. Project 3, Cu Homeostasis and Cu-ATPases in Polarized Epithelia, PI: A. Hubbard. The routes of trafficking, regulation and mechanisms involved in ATPase 7A (Menkes protein) and ATPase 7B (Wilson' protein) mediated Cu efflux from polarized epithelial cells will be studied. Associated binding proteins and the role of the C-terminus in plasma membrane targeting will be examined. Increased understanding of regulation of trafficking of plasma membrane proteins in polarized epithelia will result from these studies as well as increased understanding of the diseases which occur due to abnormal function of the transport proteins studied in each Project (diarrhea, Dent's disease, Wilson's disease, Menkes disease).

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Program Projects (P01)
Project #
1P01DK072084-01A1
Application #
7133532
Study Section
Special Emphasis Panel (ZDK1-GRB-S (M2))
Project Start
2006-09-15
Project End
2009-08-31
Budget Start
2006-09-15
Budget End
2007-08-31
Support Year
1
Fiscal Year
2006
Total Cost
$106,497
Indirect Cost
Name
Johns Hopkins University
Department
Type
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Singh, Varsha; Yang, Jianbo; Yin, Jianyi et al. (2018) Cholera toxin inhibits SNX27-retromer-mediated delivery of cargo proteins to the plasma membrane. J Cell Sci 131:
Yin, Jianyi; Tse, Chung-Ming; Avula, Leela Rani et al. (2018) Molecular Basis and Differentiation-Associated Alterations of Anion Secretion in Human Duodenal Enteroid Monolayers. Cell Mol Gastroenterol Hepatol 5:591-609
Sarker, Rafiquel; Cha, Boyoung; Kovbasnjuk, Olga et al. (2017) Phosphorylation of NHE3-S719 regulates NHE3 activity through the formation of multiple signaling complexes. Mol Biol Cell 28:1754-1767
Qiang, Xiaoling; Liotta, Anthony S; Shiloach, Joseph et al. (2017) New melanocortin-like peptide of E. coli can suppress inflammation via the mammalian melanocortin-1 receptor (MC1R): possible endocrine-like function for microbes of the gut. NPJ Biofilms Microbiomes 3:31
Cha, Boyoung; Yang, Jianbo; Singh, Varsha et al. (2017) PDZ domain-dependent regulation of NHE3 protein by both internal Class II and C-terminal Class I PDZ-binding motifs. J Biol Chem 292:8279-8290
Yu, Huimin; Hasan, Nesrin M; In, Julie G et al. (2017) The Contributions of Human Mini-Intestines to the Study of Intestinal Physiology and Pathophysiology. Annu Rev Physiol 79:291-312
Cebotaru, Liudmila; Liu, Qiangni; Yanda, Murali K et al. (2016) Inhibition of histone deacetylase 6 activity reduces cyst growth in polycystic kidney disease. Kidney Int 90:90-9
In, Julie; Foulke-Abel, Jennifer; Zachos, Nicholas C et al. (2016) Enterohemorrhagic Escherichia coli reduce mucus and intermicrovillar bridges in human stem cell-derived colonoids. Cell Mol Gastroenterol Hepatol 2:48-62.e3
Foulke-Abel, Jennifer; In, Julie; Yin, Jianyi et al. (2016) Human Enteroids as a Model of Upper Small Intestinal Ion Transport Physiology and Pathophysiology. Gastroenterology 150:638-649.e8
Gupta, Arnab; Schell, Michael J; Bhattacharjee, Ashima et al. (2016) Myosin Vb mediates Cu+ export in polarized hepatocytes. J Cell Sci 129:1179-89

Showing the most recent 10 out of 67 publications