Retinitis pigmentosa‑associated rhodopsin mutant T17M induces endoplasmic reticulum (ER) stress and sensitizes cells to ER stress-induced cell death

Jiang,Haibo; Xiong,Siqi; Xia,Xiaobo

doi:10.3892/mmr.2014.1987

Retinitis pigmentosa‑associated rhodopsin mutant T17M induces endoplasmic reticulum (ER) stress and sensitizes cells to ER stress-induced cell death

Authors:
- Haibo Jiang
- Siqi Xiong
- Xiaobo Xia
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Affiliations: Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan 410078, P.R. China
Published online on: February 26, 2014 https://doi.org/10.3892/mmr.2014.1987
Pages: 1737-1742

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Abstract

Retinitis pigmentosa (RP) is a group of inherited diseases that primarily affect light‑sensitive rods and cones in the retina. Rhodopsin mutations, including the T17M mutation, are associated with the autosomal dominant form of retinitis pigmentosa (ADRP) and have been linked to abnormal protein folding. However, the molecular mechanisms underlying T17M rhodopsin‑induced retinal degeneration are yet to be elucidated. In the present study, Human embryonic kidney (HEK) 293 and ARPE‑19 cells were transfected with myc‑tagged wild‑type (WT) and T17M rhodopsin constructs. Cells were fixed and stained with anti‑myc antibodies and the localization of WT and T17M rhodopsin was visualized using immunofluorescence microscopy. Turnover rates of WT and T17M rhodopsin were measured using western blot analysis. In addition, endoplasmic reticulum (ER) stress‑induced cell death was analyzed in WT and T17M rhodopsin‑transfected cells using nuclear staining. Misfolded T17M rhodopsin was observed to be abnormally localized in the ER, while WT rhodopsin was predominantly found at the plasma membrane. Protein turnover analysis revealed that T17M rhodopsin was more rapidly degraded by proteasomes than WT rhodopsin. Furthermore, overexpression of T17M rhodopsin was observed to induce cell death and increase cytotoxicity; predisposing cells to ER stress‑induced cell death. These findings show novel insight into the properties of T17M rhodopsin and highlight the role of ER stress in T17M‑associated RP.

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1	Dryja TP, McGee TL, Hahn LB, et al: Mutations within the rhodopsin gene in patients with autosomal dominant retinitis pigmentosa. N Engl J Med. 323:1302–1307. 1990. View Article : Google Scholar : PubMed/NCBI
2	Hartong DT, Berson EL and Dryja TP: Retinitis pigmentosa. Lancet. 368:1795–1809. 2006. View Article : Google Scholar : PubMed/NCBI
3	Mendes HF, van der Spuy J, Chapple JP and Cheetham ME: Mechanisms of cell death in rhodopsin retinitis pigmentosa: implications for therapy. Trends Mol Med. 11:177–185. 2005. View Article : Google Scholar : PubMed/NCBI
4	Nathans J and Hogness DS: Isolation and nucleotide sequence of the gene encoding human rhodopsin. Proc Natl Acad Sci USA. 81:4851–4855. 1984. View Article : Google Scholar : PubMed/NCBI
5	Kaushal S and Khorana HG: Structure and function in rhodopsin 7. Point mutations associated with autosomal dominant retinitis pigmentosa. Biochemistry. 33:6121–6128. 1994. View Article : Google Scholar : PubMed/NCBI
6	Sung CH, Davenport CM and Nathans J: Rhodopsin mutations responsible for autosomal dominant retinitis pigmentosa. Clustering of functional classes along the polypeptide chain. J Biol Chem. 268:26645–26649. 1993.
7	Li T, Sandberg MA, Pawlyk BS, et al: Effect of vitamin A supplementation on rhodopsin mutants threonine-17 --> methionine and proline-347 --> serine in transgenic mice and in cell cultures. Proc Natl Acad Sci USA. 95:11933–11938. 1998. View Article : Google Scholar : PubMed/NCBI
8	Jacobson SG, Kemp CM, Sung CH and Nathans J: Retinal function and rhodopsin levels in autosomal dominant retinitis pigmentosa with rhodopsin mutations. Am J Ophthalmol. 112:256–271. 1991. View Article : Google Scholar : PubMed/NCBI
9	Sung CH, Davenport CM, Hennessey JC, et al: Rhodopsin mutations in autosomal dominant retinitis pigmentosa. Proc Natl Acad Sci USA. 88:6481–6485. 1991. View Article : Google Scholar : PubMed/NCBI
10	White DA, Fritz JJ, Hauswirth WW, Kaushal S and Lewin AS: Increased sensitivity to light-induced damage in a mouse model of autosomal dominant retinal disease. Invest Ophthalmol Vis Sci. 48:1942–1951. 2007. View Article : Google Scholar : PubMed/NCBI
11	Ridge KD, Abdulaev NG, Sousa M and Palczewski K: Phototransduction: crystal clear. Trends Biochem Sci. 28:479–487. 2003. View Article : Google Scholar
12	Chapple JP, Grayson C, Hardcastle AJ, Saliba RS, van der Spuy J and Cheetham ME: Unfolding retinal dystrophies: a role for molecular chaperones? Trends Mol Med. 7:414–421. 2001. View Article : Google Scholar : PubMed/NCBI
13	Gorbatyuk MS, Knox T, LaVail MM, et al: Restoration of visual function in P23H rhodopsin transgenic rats by gene delivery of BiP/Grp78. Proc Natl Acad Sci USA. 107:5961–5966. 2010. View Article : Google Scholar : PubMed/NCBI
14	Krebs MP, White DA and Kaushal S: Biphasic photoreceptor degeneration induced by light in a T17M rhodopsin mouse model of cone bystander damage. Invest Ophthalmol Vis Sci. 50:2956–2965. 2009. View Article : Google Scholar : PubMed/NCBI
15	Krebs MP, Holden DC, Joshi P, Clark CL III, Lee AH and Kaushal S: Molecular mechanisms of rhodopsin retinitis pigmentosa and the efficacy of pharmacological rescue. J Mol Biol. 395:1063–1078. 2010. View Article : Google Scholar : PubMed/NCBI
16	Tam BM and Moritz OL: The role of rhodopsin glycosylation in protein folding, trafficking, and light-sensitive retinal degeneration. J Neurosci. 29:15145–15154. 2009. View Article : Google Scholar : PubMed/NCBI
17	Selkoe DJ: Cell biology of protein misfolding: The examples of Alzheimer’s and Parkinson’s diseases. Nat Cell Biol. 6:1054–1061. 2004.
18	Wickner S, Maurizi MR and Gottesman S: Posttranslational quality control: folding, refolding, and degrading proteins. Science. 286:1888–1893. 1999. View Article : Google Scholar : PubMed/NCBI
19	Chaudhuri TK and Paul S: Protein-misfolding diseases and chaperone-based therapeutic approaches. FEBS J. 273:1331–1349. 2006. View Article : Google Scholar : PubMed/NCBI
20	Kunte MM, Choudhury S, Manheim JF, et al: ER stress is involved in T17M rhodopsin-induced retinal degeneration. Invest Ophthalmol Vis Sci. 53:3792–3800. 2012. View Article : Google Scholar : PubMed/NCBI
21	Finger A, Knop M and Wolf DH: Analysis of two mutated vacuolar proteins reveals a degradation pathway in the endoplasmic reticulum or a related compartment of yeast. Eur J Biochem. 218:565–574. 1993. View Article : Google Scholar : PubMed/NCBI
22	Zhao L and Ackerman SL: Endoplasmic reticulum stress in health and disease. Curr Opin Cell Biol. 18:444–452. 2006. View Article : Google Scholar
23	Griciuc A, Aron L and Ueffing M: ER stress in retinal degeneration: a target for rational therapy? Trends Mol Med. 17:442–451. 2011. View Article : Google Scholar : PubMed/NCBI
24	Scorrano L, Oakes SA, Opferman JT, et al: BAX and BAK regulation of endoplasmic reticulum Ca²⁺: a control point for apoptosis. Science. 300:135–139. 2003. View Article : Google Scholar : PubMed/NCBI
25	Haynes CM, Titus EA and Cooper AA: Degradation of misfolded proteins prevents ER-derived oxidative stress and cell death. Mol Cell. 15:767–776. 2004. View Article : Google Scholar : PubMed/NCBI
26	Stefani IC, Wright D, Polizzi KM and Kontoravdi C: The role of ER stress-induced apoptosis in neurodegeneration. Curr Alzheimer Res. 9:373–387. 2012. View Article : Google Scholar : PubMed/NCBI
27	Hayashi T, Saito A, Okuno S, Ferrand-Drake M, Dodd RL and Chan PH: Damage to the endoplasmic reticulum and activation of apoptotic machinery by oxidative stress in ischemic neurons. J Cereb Blood Flow Metab. 25:41–53. 2005. View Article : Google Scholar : PubMed/NCBI