Journal
JOURNAL OF CELLULAR BIOCHEMISTRY
Volume 99, Issue 2, Pages 647-664Publisher
WILEY
DOI: 10.1002/jcb.20992
Keywords
scribble; lethal giant larvae; Vangl2; polarity
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Funding
- NIDDK NIH HHS [DK64388, DK63096, DK07632] Funding Source: Medline
- NIGMS NIH HHS [GM064645, R01 GM054712] Funding Source: Medline
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Scribble (Scrib) is a large multi-domain cytoplasmic protein that was first identified through its requirement for the establishment of epithelial polarity. We tested the hypotheses that Scrib asssociates with the basolateral membrane via multiple domains, binds specific protein partners, and is part of a multimeric complex. We generated a series of EGFP-tagged Scrib fusion proteins and examined their membrane localizations in two types of polarized mammalian epithelial cells using biochemical and morphological approaches. We found that Scrib's (L) under bar eucine-(r) under bar ich-(r) under bar epeat (LRR) and PDS-95/Discs Large/ZO-1 (PDZ) domains independently associate with the plasma membrane in both cell types. We identified multiple large Scrib complexes, demonstrated that Scrib and the cytoplasmic protein Lethal giant larvae2 (Lgl2) co-IP and that this association occurs via Scrib's LRR domain. Further, this report demonstrates that the membrane protein Vangl2 binds selectively to specific PDZ domains in Scrib. Our identification of Scrib's associations highlights its function in multiple biologic pathways and sets the stage for future identification of more proteins that must interact with Scrib's remaining domains. J. Cell. Biochem. 99: 647-664, 2006.(c) 2006 Wiley-Liss, Inc.
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