September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
RPGR, a retinal ciliopathy protein, localizes to cilia in a prenylation dependent manner
Author Affiliations & Notes
  • Wei Zhang
    Ophthalmology, University of Massachusetts Medical School, Worcester, Massachusetts, United States
  • Nageswara Rao Kollu
    Ophthalmology, University of Massachusetts Medical School, Worcester, Massachusetts, United States
  • Linjing Li
    Ophthalmology, University of Massachusetts Medical School, Worcester, Massachusetts, United States
  • Manisha Anand
    Ophthalmology, University of Massachusetts Medical School, Worcester, Massachusetts, United States
  • Hemant Khanna
    Ophthalmology, University of Massachusetts Medical School, Worcester, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Wei Zhang, None; Nageswara Kollu, None; Linjing Li, None; Manisha Anand, None; Hemant Khanna, None
  • Footnotes
    Support  National Eye Institute (EY022372);University of Massachusetts Center for Clinical and Translational Sciences (UMCCTS); Foundation Fighting Blindness; UMASS Cell Biology Confocal Core and Electron Microscopy Core (Award # S10RR027897)
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 3193. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Wei Zhang, Nageswara Rao Kollu, Linjing Li, Manisha Anand, Hemant Khanna; RPGR, a retinal ciliopathy protein, localizes to cilia in a prenylation dependent manner. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3193.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : RPGR (retinitis pigmentosa GTPase regulator), a major cause of retinitis pigmentosa (RP) worldwide, is a ciliary protein involved in regulating protein trafficking and composition of cilia. We undertook this study to understand the mechanisms of localization of RPGR to the cilium.

Methods : N-terminal GFP or SBP-tagged full-length and mutant human RPGR constructs were transiently transfected into hTERT-RPE1 cells grown on glass cover slips. RPGR cellular localization was detected by anti-GFP antibody; cilia were stained with anti-ARL13B antibody. RPGR-interacting proteins were identified using Tandem Affinity Purification of SBP-RPGR complexes from HEK293 cells followed by mass spectrometry analysis. Co-immunoprecipitation was performed to ascertain the interaction of RPGR and identified proteins.

Results : The C-terminal isoprenylation motif (-812CTIL815) of RPGR is essential for its localization to cilia. We found that this motif interacts with PDE6D (delta subunit of Phosphodiesterase), a prenyl-binding protein. Disruption of the -CTIL motif abrogated RPGR’s interaction with PDE6D. Selected human disease-causing mutations in RPGR perturb its ciliary localization and interaction with PDE6D.

Conclusions : Our results suggest that prenylation modulates the trafficking of a regulator of ciliary composition and indicate the involvement of mistrafficking of RPGR as a potential mechanism of associated retinal ciliopathy.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×