The deregulation of the ubiquitin-proteasome system (UPS) is a hallmark feature of many neurodegenerative disorders, including retinal degenerations, which involve misfolding and deregulation of protein turnover in photoreceptors. Our long-range objective is to understand the role of a regulatory complex of the UPS- chaperone machinery in controlling substrate degradation and folding and use this information to identify targets for novel therapeutic strategies, which presently lack efficacy toward selective biological and disease processes of the UPS. The goal of this proposal is to understand the molecular and intracellular mechanisms that sustain the balance between turnover and biogenesis of proteins, two tightly coupled processes that are vital for retinal-neuron function and survival and that are implicated in the pathogenesis of a wide variety of neuro-visual disorders of the human. Achievement of this goal will allow us and others to identify novel pharmacological targets and develop therapeutic strategies to manipulate the UPS-chaperone machinery, prevent the toxic deregulation of normal or expression of mutated proteins under various retinal disease states and aging manifestations causing the impairment of the UPS-chaperone machinery and neuro-visual function. The proposed research builds upon our prior research findings which demonstrated that substrates are presented to the proteasome upon synthesis and that the activity of the proteasome is regulated and compartmentalized in the cell. Ran-binding protein 2 (RanBP2) via two distinct domains, the cyclophilin-like (CLD) and cyclophilin (CY) domains, is a key controller of such processes. This proposal tests our overall hypothesis that RanBP2 and its CLD and CY domains control two tightly coupled processes, the UPS-dependent turnover and biogenesis of selective proteins that are critical to the function and survival of selective retinal neurons and retinal pigment epithelium (RPE) cells under normal and pathophysiological conditions. We will test this hypothesis by accomplishing the following specific aims, which focus on the mechanistic roles of RanBP2, and its CLD and CY domains, in the regulation of protein homeostasis and survival of selective retinal neurons under normal and disease stress conditions.
Aim 1. Test the hypothesis that loss-of-function of RanBP2 has differential biological and pathological effects among retinal neurons, some of which may be essential to the viability or protein homeostasis of such neurons.
Aim 2. Test the hypothesis that selective loss-of-function of the CLD activity of RanBP2 up-regulates selective functions of the UPS that are critical to homeostasis of selective retinal proteins, function and survival of retinal neurons and RPE cells.
Aim 3. Test the hypothesis that the cis-trans prolyl isomerase (PPIase) activity in the CY domain of RanBP2 controls the biogenesis of selective proteins that are critical to the function or survival of retinal neurons and RPE cells.

Public Health Relevance

The modulation of protein degradation and biogenesis in the retina is of vital importance to its function. Impairment of these processes is known to underlie the pathogenesis of several degenerative and aging disorders affecting retinal neurons. The proposal aims at elucidating the role of Ran-binding protein 2 in protein homeostasis and allied pathohysiological states in the retina.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY019492-03
Application #
8289556
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Neuhold, Lisa
Project Start
2010-08-01
Project End
2014-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
3
Fiscal Year
2012
Total Cost
$374,400
Indirect Cost
$134,400
Name
Duke University
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Patil, Hemangi; Saha, Arjun; Senda, Eugene et al. (2014) Selective impairment of a subset of Ran-GTP-binding domains of ran-binding protein 2 (Ranbp2) suffices to recapitulate the degeneration of the retinal pigment epithelium (RPE) triggered by Ranbp2 ablation. J Biol Chem 289:29767-89
Cho, Kyoung-in; Patil, Hemangi; Senda, Eugene et al. (2014) Differential loss of prolyl isomerase or chaperone activity of Ran-binding protein 2 (Ranbp2) unveils distinct physiological roles of its cyclophilin domain in proteostasis. J Biol Chem 289:4600-25
Cho, Kyoung-In; Haque, Mdemdadul; Wang, Jessica et al. (2013) Distinct and atypical intrinsic and extrinsic cell death pathways between photoreceptor cell types upon specific ablation of Ranbp2 in cone photoreceptors. PLoS Genet 9:e1003555
Patil, Hemangi; Cho, Kyoung-in; Lee, James et al. (2013) Kinesin-1 and mitochondrial motility control by discrimination of structurally equivalent but distinct subdomains in Ran-GTP-binding domains of Ran-binding protein 2. Open Biol 3:120183
Ferreira, Paulo A; Orry, Andrew (2012) From Drosophila to humans: reflections on the roles of the prolyl isomerases and chaperones, cyclophilins, in cell function and disease. J Neurogenet 26:132-43
Fernandez-Martinez, Lorena; Letteboer, Stef; Mardin, Christian Y et al. (2011) Evidence for RPGRIP1 gene as risk factor for primary open angle glaucoma. Eur J Hum Genet 19:445-51
Cho, Kyoung-in; Yi, Haiqing; Tserentsoodol, Nomingerel et al. (2010) Neuroprotection resulting from insufficiency of RANBP2 is associated with the modulation of protein and lipid homeostasis of functionally diverse but linked pathways in response to oxidative stress. Dis Model Mech 3:595-604