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

Research output: Contribution to journalArticle

122 Citations (Scopus)

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 b 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.

Original languageEnglish (US)
Pages (from-to)2347-2360
Number of pages14
JournalGenes and Development
Volume25
Issue number22
DOIs
StatePublished - Nov 15 2011

Fingerprint

Cilia
GTP Phosphohydrolases
Membranes
Myristic Acid
Proteins
Cystic Kidney Diseases
Retinal Degeneration
Neural Tube Defects
Structural Models
Guanosine Triphosphate
Phenylalanine
Proteomics
Retinitis Pigmentosa 2
Phenotype
Ciliopathies

Keywords

  • ARL3
  • Myristoylation
  • NPHP
  • UNC119

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

Cite this

Wright, K. J., Baye, L. M., Olivier-Mason, A., Mukhopadhyay, S., Sang, L., Kwong, M., ... Jackson, P. K. (2011). An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium. Genes and Development, 25(22), 2347-2360. https://doi.org/10.1101/gad.173443.111

An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium. / Wright, Kevin J.; Baye, Lisa M.; Olivier-Mason, Anique; Mukhopadhyay, Saikat; Sang, Liyun; Kwong, Mandy; Wang, Weiru; Pretorius, Pamela R.; Sheffield, Val C.; Sengupta, Piali; Slusarski, Diane C.; Jackson, Peter K.

In: Genes and Development, Vol. 25, No. 22, 15.11.2011, p. 2347-2360.

Research output: Contribution to journalArticle

Wright, KJ, Baye, LM, Olivier-Mason, A, Mukhopadhyay, S, Sang, L, Kwong, M, Wang, W, Pretorius, PR, Sheffield, VC, Sengupta, P, Slusarski, DC & Jackson, PK 2011, 'An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium', Genes and Development, vol. 25, no. 22, pp. 2347-2360. https://doi.org/10.1101/gad.173443.111
Wright, Kevin J. ; Baye, Lisa M. ; Olivier-Mason, Anique ; Mukhopadhyay, Saikat ; Sang, Liyun ; Kwong, Mandy ; Wang, Weiru ; Pretorius, Pamela R. ; Sheffield, Val C. ; Sengupta, Piali ; Slusarski, Diane C. ; Jackson, Peter K. / An ARL3-UNC119-RP2 GTPase cycle targets myristoylated NPHP3 to the primary cilium. In: Genes and Development. 2011 ; Vol. 25, No. 22. pp. 2347-2360.
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