Differential Loss of Prolyl-Isomerase or Chaperone Activity of Ran-binding protein 2 (Ranbp2) Unveils Distinct Physiological Roles of Its Cyclophilin Domain in Proteostasis [Protein Structure and Folding]

January 8th, 2014 by Cho, K.-i., Patil, H., Senda, E., Wang, J., Yi, H., Qiu, S., Yoon, D., Yu, M., Orry, A., Peachey, N. S., Ferreira, P. A.

The immunophilins, cyclophilins, catalyze peptidyl cis-trans prolyl isomerization (PPIase), a rate-limiting step in protein folding and conformational switch in protein function. Cyclophilins are also chaperones. Non-catalytic mutations affecting the only cyclophilins with known but distinct physiological substrates, the Drosophila NinaA and its mammalian homolog, cyclophilin-B, impair opsin biogenesis and cause osteogenesis imperfecta, respectively. However, the physiological roles and substrates of most cyclophilins remain unknown. It is also unclear if PPIase and chaperone activities reflect distinct cyclophilin properties. To elucidate the physiological idiosyncrasy stemming from potential cyclophilin functions, we generated mice lacking endogenous Ran-binding protein-2 (Ranbp2) and expressing bacterial artificial chrosomosomes of Ranbp2 with impaired C-terminal chaperone and with (Tg-Ranbp2WT-HA) or without PPIase activities (Tg-Ranbp2R2944A-HA). The transgenic lines exhibit unique effects in proteostasis. Either line presents selective deficits in M-opsin biogenesis with its accumulation and aggregation in cone photoreceptors, but without proteostatic impairment of two novel Ranbp2 cyclophilin partners, the cytokine-responsive effectors, STAT3/STAT5. Stress-induced STAT3 activation is also unaffected in Tg-Ranbp2R2944A-HA::Ranbp2-/-. Conversely, proteomic analyses found that the multisystem proteinopathy/ALS protein, heterogeneous nuclear ribonucleoproteins A2/B1, are down-regulated post-transcriptionally only in Tg-Ranbp2R2944A-HA::Ranbp2-/-. This is accompanied by the age and tissue-dependent reductions of diubiquitin and ubiquitylated proteins, increased deubiquitylation activity, and accumulation of the 26S proteasome subunits, S1 and S5b. These manifestations are absent in another line, Tg-Ranbp2CLDm-HA::Ranbp2-/-, harboring SUMO-1 and S1-binding mutations in the Ranbp2 cyclophilin-like domain. These results unveil distinct mechanistic and biological links between PPIase and chaperone activities of Ranbp2 cyclophilin towards proteostasis of selective substrates and with novel therapeutic potential.
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