Structural and functional plasticity of subcellular tethering, targeting and processing of RPGRIP1 by RPGR isoforms.


Mutations affecting the retinitis pigmentosa GTPase regulator-interacting protein 1 (RPGRIP1) interactome cause syndromic retinal dystrophies. RPGRIP1 interacts with the retinitis pigmentosa GTPase regulator (RPGR) through a domain homologous to RCC1 (RHD), a nucleotide exchange factor of Ran GTPase. However, functional relationships between RPGR and RPGRIP1 and their subcellular roles are lacking. We show by molecular modeling and analyses of RPGR disease-mutations that the RPGR-interacting domain (RID) of RPGRIP1 embraces multivalently the shared RHD of RPGR(1-19) and RPGR(ORF15) isoforms and the mutations are non-overlapping with the interface found between RCC1 and Ran GTPase. RPGR disease-mutations grouped into six classes based on their structural locations and differential impairment with RPGRIP1 interaction. RPGRIP1α(1) expression alone causes its profuse self-aggregation, an effect suppressed by co-expression of either RPGR isoform before and after RPGRIP1α(1) self-aggregation ensue. RPGR(1-19) localizes to the endoplasmic reticulum, whereas RPGR(ORF15) presents cytosolic distribution and they determine uniquely the subcellular co-localization of RPGRIP1α(1). Disease mutations in RPGR(1) (-19), RPGR(ORF15), or RID of RPGRIP1α(1), singly or in combination, exert distinct effects on the subcellular targeting, co-localization or tethering of RPGRIP1α(1) with RPGR(1-19) or RPGR(ORF15) in kidney, photoreceptor and hepatocyte cell lines. Additionally, RPGR(ORF15), but not RPGR(1-19), protects the RID of RPGRIP1α(1) from limited proteolysis. These studies define RPGR- and cell-type-dependent targeting pathways with structural and functional plasticity modulating the expression of mutations in RPGR and RPGRIP1. Further, RPGR isoforms distinctively determine the subcellular targeting of RPGRIP1α(1,) with deficits in RPGR(ORF15)-dependent intracellular localization of RPGRIP1α(1) contributing to pathomechanisms shared by etiologically distinct syndromic retinal dystrophies.





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Publication Info

Patil, Hemangi, Mallikarjuna R Guruju, Kyoung-In Cho, Haiqing Yi, Andrew Orry, Hyesung Kim and Paulo A Ferreira (2012). Structural and functional plasticity of subcellular tethering, targeting and processing of RPGRIP1 by RPGR isoforms. Biol Open, 1(2). pp. 140–160. 10.1242/bio.2011489 Retrieved from

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Paulo Alexandre Ferreira

Associate Professor in Ophthalmology

The long-term goal of our research program is twofold. The first is to understand the interplay between intracellular signaling, intracellular trafficking and proteostasis in health and disease; the second is to uncover molecular players and mechanisms partaking in such processes that are amenable to therapeutic intervention in a variety of disease states. Presently, our research efforts are centered on dissecting the roles of two disease-associated protein interactomes assembled by the Ran-binding protein 2 (RanBP2) and the retinitis pigmentosa GTPase regulator-interacting protein 1 (RPGRIP1) in several neuronal cell types of the retina and brain that often undergo neurodegeneration upon a multiplicity of diseases with distinct etiologies.

The RanBP2 is a large and modular 358 kDa protein scaffold, which assembles a large multifunctional complex and acts a signal integrator of molecular and subcellular signaling and trafficking pathways critical to neuronal survival or function. Mutations or deficits in RanBP2 are linked to a variety of diseases processes ranging from neurodegeneration and necrosis to stress signaling and cancer. RanBP2 modulates the assembly or disassembly of several protein complexes with apparent disparate functions and implicated in molecular processes, such as nucleocytoplasmic and microtubule-based intracellular trafficking of proteins or organelles, protein homeostasis and biogenesis, modulation of protein-protein interactions (e.g. sumoylation), and control of cell division. Interdisciplinary approaches ranging from single molecule analysis to cell-based assays and genetically modified mouse models are employed to dissect selective cell type-dependent roles of proteins modulated dynamically by RanBP2 and underlying mechanisms in healthy and disease states.

The RPGRIP1 is also a modular protein, which associates directly with molecular partners, such as the retinitis pigmentosa GTPase regulator (RPGR) and nephrocystin-4 (NPHP4). Human mutations in the genes encoding RPGRIP1, RPGR and NPHP4 lead to severe ocular-renal, syndromic and non-syndromic retinal or renal diseases. These lead ultimately to blindness, loss of kidney function or both. Emerging data from our laboratory implicate the RPGRIP1 interactome in the regulation of the tethering, targeting, exiting and/or transport of selective retinal-renal and pre-ciliary components from the endoplasmic reticulum compartment to cilia. These processes serve as molecular determinants to the formation of subcellular structures/compartments that are critical to photoreceptor or tubular kidney cell functions . Current work is directed at dissecting: i) the biological and pathological roles of components of the RPGRIP1 interactome in retinal and kidney functions; ii) the molecular, cellular and pathophysiological bases of allelic-specific mutations and genetic heterogeneity affecting components of the RPGRIP1 interactome; iii) the identification of therapeutic targets and mechanisms dependent on the functions of the RPGRIP1 assembly complex and therapeutic approaches to delay the onset or progression of degeneration of photoreceptor, tubular kidney cells or both.

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