Polarized epithelial cells line every organ and are of utmost importance for the function of the human body. Central to epithelial cell function is the establishment and maintenance of biochemically and functionally distinct membrane domains, the apical membrane facing the lumen of an organ and the basolateral membrane that is in contact with connective tissues. Both membranes are separated by tight junctions. To maintain this architecture, epithelial cells must continuously sort newly synthesized and internalized transmembrane receptors to the correct membrane domains in the biosynthetic and endocytic pathways. Our work focuses on the molecular mechanisms that ensure correct targeting to the basolateral membrane from a central sorting station, the recycling endosomes. Previously we showed that cargos destined for the basolateral membrane are recognized by the epithelial-specific clathrin adaptor complex AP-1B for incorporation into AP-1B vesicles. This grant application proposes to follow up on these findings and to investigate the molecular mechanisms of AP-1B function, membrane recruitment, and regulation of the AP-1B-dependent pathway from recycling endosomes to the basolateral membrane. We will employ genetical, biochemical, and cell biological methods to accomplish our research goals.

Public Health Relevance

The primary function of epithelial cells is to ensure the correct nutrient and waste product exchange between the body and the environment. To fulfill these different functions, the surface of epithelial cells is divided into biochemically and functionally distinct membrane domains. Our long-term goal is to understand how polarized epithelial cells establish and maintain this asymmetry with a focus on processes that lead to correct targeting of transmembrane receptors to the membrane domain that faces connective tissues and neighboring cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM070736-09
Application #
8537925
Study Section
Membrane Biology and Protein Processing (MBPP)
Program Officer
Ainsztein, Alexandra M
Project Start
2005-02-01
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
9
Fiscal Year
2013
Total Cost
$295,340
Indirect Cost
$94,668
Name
Northwestern University at Chicago
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Fölsch, Heike (2015) Analyzing the role of AP-1B in polarized sorting from recycling endosomes in epithelial cells. Methods Cell Biol 130:289-305
Fölsch, Heike (2015) Role of the epithelial cell-specific clathrin adaptor complex AP-1B in cell polarity. Cell Logist 5:e1074331
Pigati, Lucy; Kang, Richard S; Fölsch, Heike (2013) Using replication defective viruses to analyze membrane trafficking in polarized epithelial cells. Methods Cell Biol 118:125-37
Rbaibi, Youssef; Cui, Shanshan; Mo, Di et al. (2012) OCRL1 modulates cilia length in renal epithelial cells. Traffic 13:1295-305
Ang, Su Fen; Folsch, Heike (2012) The role of secretory and endocytic pathways in the maintenance of cell polarity. Essays Biochem 53:29-39
Kang, Richard S; Folsch, Heike (2011) ARH cooperates with AP-1B in the exocytosis of LDLR in polarized epithelial cells. J Cell Biol 193:51-60
Shteyn, Elina; Pigati, Lucy; Folsch, Heike (2011) Arf6 regulates AP-1B-dependent sorting in polarized epithelial cells. J Cell Biol 194:873-87
Gephart, Jonathan D; Singh, Bhuminder; Higginbotham, James N et al. (2011) Identification of a novel mono-leucine basolateral sorting motif within the cytoplasmic domain of amphiregulin. Traffic 12:1793-804
Cook, Rita Nokes; Ang, Su Fen; Kang, Richard Seung-on et al. (2011) Analyzing the function of small GTPases by microinjection of plasmids into polarized epithelial cells. J Vis Exp :
Reales, Elena; Sharma, Nikunj; Low, Seng Hui et al. (2011) Basolateral sorting of syntaxin 4 is dependent on its N-terminal domain and the AP1B clathrin adaptor, and required for the epithelial cell polarity. PLoS One 6:e21181

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