An elongated tract of polyQ in the carboxyl‑terminus of human α1A calcium channel induces cell apoptosis by nuclear translocation

Sun,Ji; Sun,Xiguang; Li,Zhuo; Ma,Dihui; Lv,Yudan

doi:10.3892/or.2020.7592

An elongated tract of polyQ in the carboxyl‑terminus of human α1A calcium channel induces cell apoptosis by nuclear translocation

Authors:
- Ji Sun
- Xiguang Sun
- Zhuo Li
- Dihui Ma
- Yudan Lv
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Affiliations: Department of Pediatric Neurology, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Hand Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Endocrinology and Metabolism, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Neurology, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: April 22, 2020 https://doi.org/10.3892/or.2020.7592
Pages: 156-164
Copyright: © Sun et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

An aberrant elongated tract of glutamine residues (polyQ) in proteins induces multiple diseases treated in the clinic. In our previous study of progressive myoclonic epilepsy (PME), using whole‑exome sequencing, a mutant Cav2.1 protein with an aberrant elongated polyQ tract was identified in PME patients. To investigate the molecular mechanism and cell biology of this aberrant elongated polyQ tract, wild‑type Cav2.1 with 13 polyQ repeats (Cav2.1 wt‑Q13) and mutant‑type Cav2.1 with 26 polyQ repeats (Cav2.1 mt‑Q26) were prepared and introduced into human SH‑SY5Y neuroblastoma cells. Using a WST‑1 assay, it was revealed that Cav2.1 mt‑Q26 markedly suppressed the proliferation of the SH‑SY5Y cells, a result not observed for the Cav2.1 wt‑Q13‑transfected cells. It was also revealed that Cav2.1 mt and its truncated molecules suppressed cell proliferation by inducing apoptosis rather than arresting the cell cycle. Further investigations indicated a nuclear translocation phenomenon associated with the Cav2.1 mt molecules. Mechanistically, it was revealed that the Cav2.1 mt molecules activated the Bcl‑2/Bax, caspase‑3 and poly ADP‑ribose polymerase (PARP) apoptotic pathways. The present study may provide new insights for interpreting the pathogenesis of PME and the relationship among polyQ, CACNA1A gene mutations and PME.

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1	Ophoff RA, Terwindt GM, Vergouwe MN, van Eijk R, Oefner PJ, Hoffman SM, Lamerdin JE, Mohrenweiser HW, Bulman DE, Ferrari M, et al: Familial hemiplegic migraine and episodic ataxia type-2 are caused by mutations in the Ca2+ channel gene CACNL1A4. Cell. 87:543–552. 1996. View Article : Google Scholar : PubMed/NCBI
2	Lea RA, Curtain RP, Hutchins C, Brimage PJ and Griffiths LR: Investigation of the CACNA1A gene as a candidate for typical migraine susceptibility. Am J Med Genet. 105:707–712. 2001. View Article : Google Scholar : PubMed/NCBI
3	Petrovicova A, Brozman M, Kurca E, Gobo T, Dluha J, Kalmarova K, Nosal V, Hikkelova M, Krajciova A, Burjanivova T and Sivak S: Novel missense variant of CACNA1A gene in a Slovak family with episodic ataxia type 2. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 161:107–110. 2017. View Article : Google Scholar : PubMed/NCBI
4	Bavassano C, Eigentler A, Stanika R, Obermair GJ, Boesch S, Dechant G and Nat R: Bicistronic CACNA1A gene expression in neurons derived from spinocerebellar ataxia type 6 patient-induced pluripotent stem cells. Stem Cells Dev. 26:1612–1625. 2017. View Article : Google Scholar : PubMed/NCBI
5	Khaiboullina SF, Mendelevich EG, Shigapova LH, Shagimardanova E, Gazizova G, Nikitin A, Martynova E, Davidyuk YN, Bogdanov EI, Gusev O, et al: Cerebellar atrophy and changes in cytokines associated with the CACNA1A R583Q mutation in a russian familial hemiplegic migraine type 1 family. Front Cell Neurosci. 11:2632017. View Article : Google Scholar : PubMed/NCBI
6	Grieco GS, Gagliardi S, Ricca I, Pansarasa O, Neri M, Gualandi F, Nappi G, Ferlini A and Cereda C: New CACNA1A deletions are associated to migraine phenotypes. J Headache Pain. 19:752018. View Article : Google Scholar : PubMed/NCBI
7	Lv Y, Wang Z, Liu C and Cui L: Identification of a novel CACNA1A mutation in a Chinese family with autosomal recessive progressive myoclonic epilepsy. Neuropsychiatr Dis Treat. 13:2631–2636. 2017. View Article : Google Scholar : PubMed/NCBI
8	Zhang Q, Chen ZS, An Y, Liu H, Hou Y, Li W, Lau KF, Koon AC, Ngo JCK and Chan HYE: A peptidylic inhibitor for neutralizing expanded CAG RNA-induced nucleolar stress in polyglutamine diseases. RNA. 24:486–498. 2018. View Article : Google Scholar : PubMed/NCBI
9	Roshan R, Choudhary A, Bhambri A, Bakshi B, Ghosh T and Pillai B: MicroRNA dysregulation in polyglutamine toxicity of TATA-box binding protein is mediated through STAT1 in mouse neuronal cells. J Neuroinflammation. 14:1552017. View Article : Google Scholar : PubMed/NCBI
10	von Niederhausern N, Ducray A, Zielinski J, Murbach M and Mevissen M: Effects of radiofrequency electromagnetic field exposure on neuronal differentiation and mitochondrial function in SH-SY5Y cells. Toxicol In Vitro. 61:1046092019. View Article : Google Scholar : PubMed/NCBI
11	Karikari TK, Nagel DA, Grainger A, Clarke-Bland C, Crowe J, Hill EJ and Moffat KG: Distinct conformations, aggregation and cellular internalization of different tau strains. Front Cell Neurosci. 13:2962019. View Article : Google Scholar : PubMed/NCBI
12	Liu M, Bai X, Yu S, Zhao W, Qiao J, Liu Y, Zhao D, Wang J and Wang S: Ginsenoside Re inhibits ROS/ASK-1 dependent mitochondrial apoptosis pathway and activation of Nrf2-Antioxidant response in beta-amyloid-challenged SH-SY5Y cells. Molecules. 24:E26872019. View Article : Google Scholar : PubMed/NCBI
13	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
14	Kordasiewicz HB, Thompson RM, Clark HB and Gomez CM: C-termini of P/Q-type Ca2+ channel alpha1A subunits translocate to nuclei and promote polyglutamine-mediated toxicity. Hum Mol Genet. 15:1587–1599. 2006. View Article : Google Scholar : PubMed/NCBI
15	Tsoi H, Lau TC, Tsang SY, Lau KF and Chan HY: CAG expansion induces nucleolar stress in polyglutamine diseases. Proc Natl Acad Sci USA. 109:13428–13433. 2012. View Article : Google Scholar : PubMed/NCBI
16	Mizuguchi T, Suzuki T, Abe C, Umemura A, Tokunaga K, Kawai Y, Nakamura M, Nagasaki M, Kinoshita K, Okamura Y, et al: A 12-kb structural variation in progressive myoclonic epilepsy was newly identified by long-read whole-genome sequencing. J Hum Genet. 64:359–368. 2019. View Article : Google Scholar : PubMed/NCBI
17	Tonin R, Catarzi S, Caciotti A, Procopio E, Marini C, Guerrini R and Morrone A: Progressive myoclonus epilepsy in gaucher disease due to a new Gly-Gly mutation causing loss of an exonic splicing enhancer. J Neurol. 266:92–101. 2019. View Article : Google Scholar : PubMed/NCBI
18	He J, Lin H, Li JJ, Su HZ, Wang DN, Lin Y, Wang N and Chen WJ: Identification of a novel homozygous splice-site mutation in SCARB2 that causes progressive myoclonus epilepsy with or without renal failure. Chin Med J (Engl). 131:1575–1583. 2018. View Article : Google Scholar : PubMed/NCBI
19	Algahtani H, Shirah B, Algahtani R, Al-Qahtani MH, Abdulkareem AA and Naseer MI: A novel mutation in CACNA1A gene in a Saudi female with episodic ataxia type 2 with no response to acetazolamide or 4-aminopyridine. Intractable Rare Dis Res. 8:67–71. 2019. View Article : Google Scholar : PubMed/NCBI
20	Gerhardstein BL, Gao T, Bunemann M, Puri TS, Adair A, Ma H and Hosey MM: Proteolytic processing of the C terminus of the alpha(1C) subunit of L-type calcium channels and the role of a proline-rich domain in membrane tethering of proteolytic fragments. J Biol Chem. 275:8556–8563. 2000. View Article : Google Scholar : PubMed/NCBI
21	Gomez-Ospina N, Tsuruta F, Barreto-Chang O, Hu L and Dolmetsch R: The C terminus of the L-type voltage-gated calcium channel Ca(V)1.2 encodes a transcription factor. Cell. 127:591–606. 2006. View Article : Google Scholar : PubMed/NCBI
22	Park D, Kim SH, Lee YJ, Song GJ and Park JS: A novel CACNA1A mutation associated with episodic ataxia 2 presenting with periodic paralysis. Acta Neurol Belg. 118:137–139. 2018. View Article : Google Scholar : PubMed/NCBI
23	Walker FO: Huntington's disease. Lancet. 369:218–228. 2007. View Article : Google Scholar : PubMed/NCBI
24	Miyata R, Hayashi M, Tanuma N, Shioda K, Fukatsu R and Mizutani S: Oxidative stress in neurodegeneration in dentatorubral-pallidoluysian atrophy. J Neurol Sci. 264:133–139. 2008. View Article : Google Scholar : PubMed/NCBI
25	Sanchez I, Xu CJ, Juo P, Kakizaka A, Blenis J and Yuan J: Caspase-8 is required for cell death induced by expanded polyglutamine repeats. Neuron. 22:623–633. 1999. View Article : Google Scholar : PubMed/NCBI
26	Tien CL, Wen FC and Hsieh M: The polyglutamine-expanded protein ataxin-3 decreases bcl-2 mRNA stability. Biochem Biophys Res Commun. 365:232–238. 2008. View Article : Google Scholar : PubMed/NCBI
27	Maiuri T, Suart CE, Hung CLK, Graham KJ, Barba Bazan CA and Truant R: DNA damage repair in huntington's disease and other neurodegenerative diseases. Neurotherapeutics. 16:948–956. 2019. View Article : Google Scholar : PubMed/NCBI
28	Hwang JY, Park JH, Kim MJ, Kim WJ, Ha KT, Choi BT, Lee SY and Shin HK: Isolinderalactone regulates the BCL-2/caspase-3/PARP pathway and suppresses tumor growth in a human glioblastoma multiforme xenograft mouse model. Cancer Lett. 443:25–33. 2019. View Article : Google Scholar : PubMed/NCBI