An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium

Kevin J Wright; Lisa M Baye; Anique Olivier-Mason; Saikat Mukhopadhyay; Liyun Sang; Mandy Kwong; Weiru Wang; Pamela R Pretorius; Val C Sheffield; Piali Sengupta; Diane C Slusarski; Peter K Jackson

doi:10.1101/gad.173443.111

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An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium

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An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium

Kevin J Wright, Lisa M Baye, Anique Olivier-Mason, Saikat Mukhopadhyay, Liyun Sang, Mandy Kwong, Weiru Wang, Pamela R Pretorius, Val C Sheffield, Piali Sengupta, …

Genes & development, Vol.25(22), pp.2347-2360

11/15/2011

DOI: 10.1101/gad.173443.111

PMCID: PMC3222901

PMID: 22085962

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Abstract

The membrane of the primary cilium is a highly specialized compartment that organizes proteins to achieve spatially ordered signaling. Disrupting ciliary organization leads to diseases called ciliopathies, with phenotypes ranging from retinal degeneration and cystic kidneys to neural tube defects. How proteins are selectively transported to and organized in the primary cilium remains unclear. Using a proteomic approach, we identified the ARL3 effector UNC119 as a binding partner of the myristoylated ciliopathy protein nephrocystin-3 (NPHP3). We mapped UNC119 binding to the N-terminal 200 residues of NPHP3 and found the interaction requires myristoylation. Creating directed mutants predicted from a structural model of the UNC119-myristate complex, we identified highly conserved phenylalanines within a hydrophobic β sandwich to be essential for myristate binding. Furthermore, we found that binding of ARL3-GTP serves to release myristoylated cargo from UNC119. Finally, we showed that ARL3, UNC119b (but not UNC119a), and the ARL3 GAP Retinitis Pigmentosa 2 (RP2) are required for NPHP3 ciliary targeting and that targeting requires UNC119b myristoyl-binding activity. Our results uncover a selective, membrane targeting GTPase cycle that delivers myristoylated proteins to the ciliary membrane and suggest that other myristoylated proteins may be similarly targeted to specialized membrane domains.

Caenorhabditis elegans Proteins - chemistry

Caenorhabditis elegans Proteins - metabolism

Intracellular Signaling Peptides and Proteins - metabolism

Zebrafish - embryology

Gene Knockdown Techniques

Kinesin - genetics

Cilia - enzymology

Intracellular Signaling Peptides and Proteins - genetics

Protein Structure, Tertiary

Amino Acid Sequence

Cell Line

Caenorhabditis elegans - metabolism

Caenorhabditis elegans - genetics

Zebrafish Proteins - metabolism

Monomeric GTP-Binding Proteins - genetics

Cilia - metabolism

Zebrafish - genetics

Kinesin - metabolism

Protein Transport

Phenotype

Sequence Alignment

Animals

GTP Phosphohydrolases - metabolism

Monomeric GTP-Binding Proteins - metabolism

GTP Phosphohydrolases - genetics

Zebrafish - metabolism

Adaptor Proteins, Signal Transducing - genetics

Protein Binding

Mutation

Zebrafish Proteins - genetics

Adaptor Proteins, Signal Transducing - metabolism

Caenorhabditis elegans - enzymology

Details

Title: Subtitle: An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium
Creators: Kevin J Wright - Genentech Inc., South San Francisco, California 94080, USA
Lisa M Baye
Anique Olivier-Mason
Saikat Mukhopadhyay
Liyun Sang
Mandy Kwong
Weiru Wang
Pamela R Pretorius
Val C Sheffield
Piali Sengupta
Diane C Slusarski
Peter K Jackson
Resource Type: Journal article
Publication Details: Genes & development, Vol.25(22), pp.2347-2360
Publisher: United States
DOI: 10.1101/gad.173443.111
PMID: 22085962
PMCID: PMC3222901
ISSN: 0890-9369
eISSN: 1549-5477
Grant note: F31 DC010090 / NIDCD NIH HHS R01EY110298 / NEI NIH HHS T32 HL007121 / NHLBI NIH HHS Howard Hughes Medical Institute R37 GM56223 / NIGMS NIH HHS R01 EY017168 / NEI NIH HHS DC010090 / NIDCD NIH HHS R01EY017168 / NEI NIH HHS R01CA112369 / NCI NIH HHS R37 GM056223 / NIGMS NIH HHS R01 GM056223 / NIGMS NIH HHS R01 CA112369 / NCI NIH HHS
Language: English
Date published: 11/15/2011
Academic Unit: Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Biology; Ophthalmology and Visual Sciences
Record Identifier: 9983991962202771

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