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PMID: 18256276 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Phosphatidylinositol-4-kinase type II alpha contains an AP-3-sorting motif and a kinase domain that are both required for endosome traffic.

Molecular biology of the cell ·Vol. 19 ·No. 4 ·2008-04-00 ·Pages 1415-26

Craige B, Salazar G, Faundez V

Abstract

The adaptor complex 3 (AP-3) targets membrane proteins from endosomes to lysosomes, lysosome-related organelles and synaptic vesicles. Phosphatidylinositol-4-kinase type II alpha (PI4KIIalpha) is one of several proteins possessing catalytic domains that regulate AP-3-dependent sorting. Here we present evidence that PI4KIIalpha uniquely behaves both as a membrane protein cargo as well as an enzymatic regulator of adaptor function. In fact, AP-3 and PI4KIIalpha form a complex that requires a dileucine-sorting motif present in PI4KIIalpha. Mutagenesis of either the PI4KIIalpha-sorting motif or its kinase-active site indicates that both are necessary to interact with AP-3 and properly localize PI4KIIalpha to LAMP-1-positive endosomes. Similarly, both the kinase activity and the sorting signal present in PI4KIIalpha are necessary to rescue endosomal PI4KIIalpha siRNA-induced mutant phenotypes. We propose a mechanism whereby adaptors use canonical sorting motifs to selectively recruit a regulatory enzymatic activity to restricted membrane domains.

MeSH Terms
Amino Acid Motifs Animals Base Sequence Binding Sites/genetics Catalytic Domain/genetics Cell Line DNA-Binding Proteins/chemistry,genetics,metabolism Endosomes/metabolism Humans Lysosome-Associated Membrane Glycoproteins/metabolism Minor Histocompatibility Antigens Multiprotein Complexes Mutagenesis, Site-Directed PC12 Cells Phosphotransferases (Alcohol Group Acceptor)/antagonists & inhibitors,chemistry,genetics,metabolism Protein Structure, Tertiary RNA, Small Interfering/genetics Rats Recombinant Fusion Proteins/chemistry,genetics,metabolism Transcription Factors/chemistry,genetics,metabolism Transfection
Chemicals
DNA-Binding Proteins LAMP1 protein, human Lysosome-Associated Membrane Glycoproteins Minor Histocompatibility Antigens Multiprotein Complexes RNA, Small Interfering Recombinant Fusion Proteins Transcription Factors enhancer-binding protein AP-3 Phosphotransferases (Alcohol Group Acceptor) phosphatidylinositol phosphate 4-kinase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Craige Branch
Graduate Program in Biochemistry, Cell, and Developmental Biology, Emory University, Atlanta, GA 30322, USA.
Salazar Gloria
Faundez Victor
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.e07-12-1239
Published
2008-04-00
Epub
2008-00-06
Pages
1415-26
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2291421
Subset
IM
Grants
NINDS NIH HHS · NS42599 · United States
NINDS NIH HHS · R01 NS042599 · United States
NIGMS NIH HHS · R01 GM077569 · United States
NINDS NIH HHS · R56 NS042599 · United States
NIGMS NIH HHS · GM 077569 · United States
External Links
PubMed source DOI: 10.1091/mbc.e07-12-1239 PMC Full Text View journal details
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