Proper apico-basal epithelial polarity is crucial for normal kidney function and is perturbed in kidney diseases such as acute tubular necrosis and polycystic kidney disease. Recent advances by our group and others have provided new insights into the role of multi-protein complexes in epithelial cell polarization and protein targeting. In the current funding period our laboratory identified scaffolding proteins that localize to the tight junction and are crucial for cell polarization. Our work has focused on proteins associated with the small PDZ domain protein, mLin-7. We demonstrated that one of these mLin-7 binding partners called Protein Associated with Lin Seven 1 (PALS1) localizes to tight junctions and complexes with PALS1 Associated Tight Junction Protein (PATJ) and CrumbsS. PALS1 and PATJ are scaffolding proteins that contain PDZ and L27 domains while CrumbsS is a small apical transmembrane protein. Using siRNA and dominant negative proteins we have demonstrated that PALS1 and PATJ are crucial for epithelial cell polarity. We have also found that PALS1 directly interacts with a common polarity cassette consisting of three proteins Par3/Par6/atypical Protein Kinase C. The goal of our research is to understand how PALS1 and PATJ control the initial steps in epithelial cell polarization. We hypothesize that the scaffolding proteins, PALS1 and PATJ move from an intracellular location to mark a spot in polarizing epithelial cells that directs tight junction localization and the transition from apical to basolateral membrane surfaces. The goals of this proposal are to understand how these proteins come to identify this spot in polarizing epithelial cells and how they proceed to recruit other proteins involved in cell polarization. To achieve these goals we will examine the domains of PALS1 AND PATJ that control their movement within MDCK cells. We will also examine which domains of these proteins are important in localization, trafficking, tight junction formation and cell polarization. In addition, we will examine the signal transduction processes that control these events. These studies will shed new light on the processes necessary for early epithelial polarization and have important implication for renal epithelial function in health and disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK058208-05
Application #
6979850
Study Section
Cellular and Molecular Biology of the Kidney Study Section (CMBK)
Program Officer
Mullins, Christopher V
Project Start
2000-07-01
Project End
2010-06-30
Budget Start
2005-09-01
Budget End
2006-06-30
Support Year
5
Fiscal Year
2005
Total Cost
$317,666
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Harder, Jennifer L; Whiteman, Eileen L; Pieczynski, Jay N et al. (2012) Snail destabilizes cell surface Crumbs3a. Traffic 13:1170-85
Pieczynski, Jay; Margolis, Ben (2011) Protein complexes that control renal epithelial polarity. Am J Physiol Renal Physiol 300:F589-601
Teoh, Kim-Tat; Siu, Yu-Lam; Chan, Wing-Lim et al. (2010) The SARS coronavirus E protein interacts with PALS1 and alters tight junction formation and epithelial morphogenesis. Mol Biol Cell 21:3838-52
Schluter, Marc A; Margolis, Ben (2009) Apical lumen formation in renal epithelia. J Am Soc Nephrol 20:1444-52
Schl├╝ter, Marc A; Pfarr, Catherine S; Pieczynski, Jay et al. (2009) Trafficking of Crumbs3 during cytokinesis is crucial for lumen formation. Mol Biol Cell 20:4652-63
Whiteman, E L; Liu, C-J; Fearon, E R et al. (2008) The transcription factor snail represses Crumbs3 expression and disrupts apico-basal polarity complexes. Oncogene 27:3875-9
Shin, Kunyoo; Wang, Qian; Margolis, Ben (2007) PATJ regulates directional migration of mammalian epithelial cells. EMBO Rep 8:158-64
Wang, Qian; Chen, Xiao-Wei; Margolis, Ben (2007) PALS1 regulates E-cadherin trafficking in mammalian epithelial cells. Mol Biol Cell 18:874-85
Straight, Samuel W; Pieczynski, Jay N; Whiteman, Eileen L et al. (2006) Mammalian lin-7 stabilizes polarity protein complexes. J Biol Chem 281:37738-47
Latorre, Isabel J; Roh, Michael H; Frese, Kristopher K et al. (2005) Viral oncoprotein-induced mislocalization of select PDZ proteins disrupts tight junctions and causes polarity defects in epithelial cells. J Cell Sci 118:4283-93

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