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PMID: 16775013 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

MARK2/EMK1/Par-1Balpha phosphorylation of Rab11-family interacting protein 2 is necessary for the timely establishment of polarity in Madin-Darby canine kidney cells.

Molecular biology of the cell ·Vol. 17 ·No. 8 ·2006-08-00 ·Pages 3625-37

Ducharme NA, Hales CM, Lapierre LA, Ham AJ, Oztan A, Apodaca G, Goldenring JR

Abstract

Rab11a, myosin Vb, and the Rab11-family interacting protein 2 (FIP2) regulate plasma membrane recycling in epithelial cells. This study sought to characterize more fully Rab11-FIP2 function by identifying kinase activities modifying Rab11-FIP2. We have found that gastric microsomal membrane extracts phosphorylate Rab11-FIP2 on serine 227. We identified the kinase that phosphorylated Rab11-FIP2 as MARK2/EMK1/Par-1Balpha (MARK2), and recombinant MARK2 phosphorylated Rab11-FIP2 only on serine 227. We created stable Madin-Darby canine kidney (MDCK) cell lines expressing enhanced green fluorescent protein-Rab11-FIP2 wild type or a nonphosphorylatable mutant [Rab11-FIP2(S227A)]. Analysis of these cell lines demonstrates a new role for Rab11-FIP2 in addition to that in the plasma membrane recycling system. In calcium switch assays, cells expressing Rab11-FIP2(S227A) showed a defect in the timely reestablishment of p120-containing junctional complexes. However, Rab11-FIP2(S227A) did not affect localization with recycling system components or the normal function of apical recycling and transcytosis pathways. These results indicate that phosphorylation of Rab11-FIP2 on serine 227 by MARK2 regulates an alternative pathway modulating the establishment of epithelial polarity.

MeSH Terms
Adherens Junctions/metabolism Amino Acid Sequence Animals Calcium Signaling Cell Polarity Cells, Cultured Dogs Epithelial Cells/cytology Gene Expression Molecular Sequence Data Mutation/genetics Myosin Type V/metabolism Phosphorylation Protein Binding Protein Serine-Threonine Kinases/metabolism Protein Transport Rabbits Receptor, PAR-1/metabolism rab GTP-Binding Proteins/metabolism
Chemicals
Receptor, PAR-1 EMK1 protein, Canis familiaris Protein Serine-Threonine Kinases Myosin Type V rab11 protein rab GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Ducharme Nicole A
Department of Surgery and Cell and Developmental Biology, Vanderbilt University School of Medicine, Vanderbilt-Ingram Cancer Center, and the Nashville VA Medical Center, Nashville, TN 37232, USA.
Hales Chadwick M
Lapierre Lynne A
Ham Amy-Joan L
Oztan Asli
Apodaca Gerard
Goldenring James R
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-08-0736
Published
2006-08-00
Epub
2006-00-14
Pages
3625-37
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1525241
Subset
IM
Grants
NCI NIH HHS · CA-68485 · United States
NIDDK NIH HHS · DK-58404 · United States
NIDDK NIH HHS · R01 DK048370 · United States
NIDDK NIH HHS · DK-43405 · United States
NIDDK NIH HHS · R01 DK051970 · United States
NIDDK NIH HHS · P60 DK020593 · United States
NIDDK NIH HHS · R01 DK043405 · United States
NIDDK NIH HHS · P30 DK058404 · United States
NIDDK NIH HHS · DK-20593 · United States
NICHD NIH HHS · P30 HD015052 · United States
NIDDK NIH HHS · P30 DK020593 · United States
NCI NIH HHS · P30 CA068485 · United States
NIDDK NIH HHS · DK-48370 · United States
NICHD NIH HHS · HD15052 · United States
NIDDK NIH HHS · R01 DK-51970 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-08-0736 PMC Full Text View journal details
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