Binary Function of ARL3-GTP Revealed by Gene Knockouts

  • Christin Hanke-GogokhiaEmail author
  • Jeanne M. Frederick
  • Houbin Zhang
  • Wolfgang BaehrEmail author
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1074)


UNC119 and PDEδ are lipid-binding proteins and are thought to form diffusible complexes with transducin-α and prenylated OS proteins, respectively, to mediate their trafficking to photoreceptor outer segments. Here, we investigate mechanisms of trafficking which are controlled by Arf-like protein 3 (Arl3), a small GTPase. The activity of ARL3 is regulated by a GEF (ARL13b) and a GAP (RP2). In a mouse germline knockout of RP2, ARL3-GTP is abundant as its intrinsic GTPase activity is extremely low. High levels of ARL3-GTP impair binding and trafficking of cargo to the outer segment. Germline knockout of ARL3 is embryonically lethal generating a syndromic ciliopathy-like phenotype. Retina- and rod-specific knockout of ARL3 allow to determine the precise mechanisms leading to photoreceptor degeneration. The knockouts reveal binary functions of ARL3-GTP as a key molecule in late-stage photoreceptor ciliogenesis and cargo displacement factor.


ARL3 ARL13b PDEδ UNC119 RP2 Photoreceptor Germline knockout Retina disease 



This research was supported by a National Eye Institute grants EY08123, EY019298 (WB), EY014800-039003 (NEI core grant), unrestricted grants to the Departments of Ophthalmology at the University of Utah from Research to Prevent Blindness (RPB; New York), the Retina Research Foundation, Houston (Alice McPherson, MD), and the Foundation for Retina Research (David Brint, MD). WB is a recipient of a Research to Prevent Blindness Senior Investigator and Nelson Trust Award.


  1. Arshavsky VY, Lamb TD, Pugh EN Jr (2002) G proteins and phototransduction. Annu Rev Physiol 64:153–187CrossRefPubMedGoogle Scholar
  2. Baehr W, Devlin MJ, Applebury ML (1979) Isolation and characterization of cGMP phosphodiesterase from bovine rod outer segments. J Biol Chem 254:11669–11677PubMedGoogle Scholar
  3. Cantagrel V, Silhavy JL, Bielas SL et al (2008) Mutations in the cilia gene ARL13B lead to the classical form of Joubert syndrome. Am J Hum Genet 83:170–179CrossRefPubMedPubMedCentralGoogle Scholar
  4. Caspary T, Larkins CE, Anderson KV (2007) The graded response to Sonic Hedgehog depends on cilia architecture. Dev Cell 12:767–778CrossRefPubMedGoogle Scholar
  5. Constantine R, Zhang H, Gerstner CD et al (2012) Uncoordinated (UNC)119: coordinating the trafficking of myristoylated proteins. Vis Res 75:26–32CrossRefPubMedPubMedCentralGoogle Scholar
  6. Duldulao NA, Lee S, Sun Z (2009) Cilia localization is essential for in vivo functions of the Joubert syndrome protein Arl13b/Scorpion. Development 136:4033–4042CrossRefPubMedPubMedCentralGoogle Scholar
  7. Fung BKK, Young JH, Yamane HK et al (1990) Subunit stoichiometry of retinal rod cGMP phosphodiesterase. Biochemistry 29:2657–2664CrossRefPubMedGoogle Scholar
  8. Gilliam JC, Chang JT, Sandoval IM et al (2012) Three-dimensional architecture of the rod sensory cilium and its disruption in retinal neurodegeneration. Cell 151:1029–1041CrossRefPubMedPubMedCentralGoogle Scholar
  9. Gotthardt K, Lokaj M, Koerner C et al (2015) A G-protein activation cascade from Arl13b to Arl3 and implications for ciliary targeting of lipidated proteins. elife 4:e11859CrossRefPubMedPubMedCentralGoogle Scholar
  10. Hanke-Gogokhia C, Wu Z, Gerstner CD et al (2016) Arf-like protein 3 (ARL3) regulates protein trafficking and ciliogenesis in mouse photoreceptors. J Biol Chem 291:7142–7155CrossRefPubMedPubMedCentralGoogle Scholar
  11. Hanzal-Bayer M, Renault L, Roversi P et al (2002) The complex of Arl2-GTP and PDE delta: from structure to function. EMBO J 21:2095–2106CrossRefPubMedPubMedCentralGoogle Scholar
  12. Jayasundera T, Branham KE, Othman M et al (2010) RP2 phenotype and pathogenetic correlations in X-linked retinitis pigmentosa. Arch Ophthalmol 128:915–923CrossRefPubMedPubMedCentralGoogle Scholar
  13. Kobayashi A, Higashide T, Hamasaki D et al (2000) HRG4 (UNC119) mutation found in cone-rod dystrophy causes retinal degeneration in a transgenic model. Invest Ophthalmol Vis Sci 41:3268–3277PubMedGoogle Scholar
  14. Kuhnel K, Veltel S, Schlichting I et al (2006) Crystal structure of the human retinitis pigmentosa 2 protein and its interaction with Arl3. Structure 14:367–378CrossRefPubMedGoogle Scholar
  15. Maeda T, Imanishi Y, Palczewski K (2003) Rhodopsin phosphorylation: 30 years later. Prog Retin Eye Res 22:417–434CrossRefPubMedGoogle Scholar
  16. Manolaridis I, Kulkarni K, Dodd RB et al (2013) Mechanism of farnesylated CAAX protein processing by the intramembrane protease Rce1. Nature 504:301–305CrossRefPubMedGoogle Scholar
  17. Palczewski K, Benovic JL (1991) G-protein-coupled receptor kinases. TIBS 16:387–391PubMedPubMedCentralGoogle Scholar
  18. Schrick JJ, Vogel P, Abuin A et al (2006) ADP-ribosylation factor-like 3 is involved in kidney and photoreceptor development. Am J Pathol 168:1288–1298CrossRefPubMedPubMedCentralGoogle Scholar
  19. Strom SP, Clark MJ, Martinez A et al (2016) De novo occurrence of a variant in ARL3 and apparent autosomal dominant transmission of retinitis pigmentosa. PLoS One 11:e0150944CrossRefPubMedPubMedCentralGoogle Scholar
  20. Thomas S, Wright KJ, Le CS et al (2014) A homozygous PDE6D mutation in Joubert syndrome impairs targeting of farnesylated INPP5E protein to the primary cilium. Hum Mutat 35:137–146CrossRefPubMedPubMedCentralGoogle Scholar
  21. Veltel S, Gasper R, Eisenacher E et al (2008) The retinitis pigmentosa 2 gene product is a GTPase-activating protein for Arf-like 3. Nat Struct Mol Biol 15:373–380CrossRefPubMedGoogle Scholar
  22. Wright AF, Chakarova CF, Abd El-Aziz MM et al (2010) Photoreceptor degeneration: genetic and mechanistic dissection of a complex trait. Nat Rev Genet 11:273–284CrossRefPubMedPubMedCentralGoogle Scholar
  23. Young RW (1976) Visual cells and the concept of renewal. Invest Ophthalmol Vis Sci 15:700–725PubMedGoogle Scholar
  24. Zhang H, Li S, Doan T et al (2007) Deletion of PrBP/{delta} impedes transport of GRK1 and PDE6 catalytic subunits to photoreceptor outer segments. Proc Natl Acad Sci U S A 104:8857–8862CrossRefPubMedPubMedCentralGoogle Scholar
  25. Zhang H, Constantine R, Vorobiev S et al (2011) UNC119 is required for G protein trafficking in sensory neurons. Nat Neurosci 14:874–880CrossRefPubMedPubMedCentralGoogle Scholar
  26. Zhang H, Constantine R, Frederick JM et al (2012) The prenyl-binding protein PrBP/delta: a chaperone participating in intracellular trafficking. Vis Res 75:19–25CrossRefPubMedPubMedCentralGoogle Scholar
  27. Zhang H, Hanke-Gogokhia C, Jiang L et al (2015) Mistrafficking of prenylated proteins causes retinitis pigmentosa 2. FASEB J 29:932–942CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Ophthalmology and Visual SciencesJohn A. Moran Eye Center, University of Utah School of MedicineSalt Lake CityUSA
  2. 2.Department of Biochemistry and BiologyUniversity of PotsdamPotsdamGermany
  3. 3.The Sichuan Provincial Key Laboratory for Human Disease Gene StudyThe Institute of Laboratory Medicine, Hospital of University of Electronic Science and Technology of China and Sichuan Provincial People’s HospitalChengduChina
  4. 4.School of Medicine, University of Electronic Science and Technology of ChinaChengduChina
  5. 5.Department of Neurobiology and AnatomyUniversity of Utah School of MedicineSalt Lake CityUSA
  6. 6.Department of BiologyUniversity of UtahSalt Lake CityUSA

Personalised recommendations