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The E3 Ligase PIAS1 Regulates P53 Sumoylation to Control Stress-Induced Apoptosis of Lens

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The E3 Ligase PIAS1 Regulates P53 Sumoylation to Control Stress-Induced Apoptosis of Lens fcell-09-660494 June 14, 2021 Time: 13:5 # 1 ORIGINAL RESEARCH published: 14 June 2021 doi: 10.3389/fcell.2021.660494 The E3 Ligase PIAS1 Regulates p53 Sumoylation to Control Stress-Induced Apoptosis of Lens Edited by: Wolfgang Knabe, Epithelial Cells Through the Universität Münster, Germany Proapoptotic Regulator Bax Reviewed by: Guillaume Bossis, † † † † † Centre National de la Recherche Qian Nie , Huimin Chen , Ming Zou , Ling Wang , Min Hou , Jia-Wen Xiang, Scientifique (CNRS), France Zhongwen Luo, Xiao-Dong Gong, Jia-Ling Fu, Yan Wang, Shu-Yu Zheng, Yuan Xiao, Stefan Müller, Yu-Wen Gan, Qian Gao, Yue-Yue Bai, Jing-Miao Wang, Lan Zhang, Xiang-Cheng Tang, Goethe University Frankfurt, Germany Xuebin Hu, Lili Gong, Yizhi Liu* and David Wan-Cheng Li*‡ Leena Latonen, University of Eastern Finland, Finland State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China *Correspondence: David Wan-Cheng Li Protein sumoylation is one of the most important post-translational modifications [email protected] Yizhi Liu regulating many biological processes (Flotho A & Melchior F. 2013. Ann Rev. Biochem. [email protected] 82:357–85). Our previous studies have shown that sumoylation plays a fundamental †These authors have contributed role in regulating lens differentiation (Yan et al., 2010. PNAS, 107(49):21034-9.; equally to this work Gong et al., 2014. PNAS. 111(15):5574–9). Whether sumoylation is implicated in lens ‡ ORCID: David Wan-Cheng Li pathogenesis remains elusive. Here, we present evidence to show that the protein orcid.org/0000-0002-7398-7630 inhibitor of activated STAT-1 (PIAS1), a E3 ligase for sumoylation, is implicated in regulating stress-induced lens pathogenesis. During oxidative stress-induced Specialty section: cataractogenesis, expression of PIAS1 is significantly altered at both mRNA and protein This article was submitted to Cell Death and Survival, levels. Upregulation and overexpression of exogenous PIAS1 significantly enhances a section of the journal stress-induced apoptosis. In contrast, silence of PIAS1 with CRISPR/Cas9 technology Frontiers in Cell and Developmental attenuates stress-induced apoptosis. Mechanistically, different from other cells, PIAS1 Biology has little effect to activate JNK but upregulates Bax, a major proapoptotic regulator. Received: 29 January 2021 Accepted: 20 April 2021 Moreover, Bax upregulation is derived from the enhanced transcription activity of the Published: 14 June 2021 upstream transcription factor, p53. As revealed previously in other cells by different Citation: laboratories, our data also demonstrate that PIAS1 promotes SUMO1 conjugation of Nie Q, Chen H, Zou M, Wang L, Hou M, Xiang J-W, Luo Z, Gong X-D, p53 at K386 residue in lens epithelial cells and thus enhances p53 transcription activity Fu J-L, Wang Y, Zheng S-Y, Xiao Y, to promote Bax upregulation. Silence of Bax expression largely abrogates PIAS1- Gan Y-W, Gao Q, Bai Y-Y, Wang J-M, mediated enhancement of stress-induced apoptosis. Thus, our results demonstrated Zhang L, Tang X-C, Hu X, Gong L, Liu Y and Li DW (2021) The E3 Ligase that PIAS1 promotes oxidative stress-induced apoptosis through positive control of p53, PIAS1 Regulates p53 Sumoylation to which specifically upregulates expression of the downstream proapoptotic regulator Control Stress-Induced Apoptosis of Lens Epithelial Cells Through Bax. As a result, PIAS1-promoted apoptosis induced by oxidative stress is implicated in the Proapoptotic Regulator Bax. lens pathogenesis. Front. Cell Dev. Biol. 9:660494. doi: 10.3389/fcell.2021.660494 Keywords: sumoylation, apoptosis, PIAS1, p53, Bax, lens, cataract, oxidative stress Frontiers in Cell and Developmental Biology| www.frontiersin.org 1 June 2021| Volume 9| Article 660494 fcell-09-660494 June 14, 2021 Time: 13:5 # 2 Nie et al. PIAS1 Regulates Apoptosis Through p53-Bax INTRODUCTION is implicated in ocular diseases, especially lens cataractogenesis, remains elusive. Sumoylation is a unique class of protein post-translational Cataract is an aging disease that, in most cases, is derived from modification, discovered in the past century (Boddy et al., aging process or stress induction such as oxidative stress (Spector, 1996; Matunis et al., 1996; Okura et al., 1996; Shen et al., 1995 and references therein). Mechanistically, we have previously 1996). The small ubiquitin-like modifier (SUMO) proteins can demonstrated that stress-induced apoptosis is a common cellular interact with over 6,000 proteins, regulating their activity, basis for non-congenital cataractogenesis (Li et al., 1995a,b; Li interactions, localization, or stability through a reversible and Spector, 1996). In lens epithelial cells (LECs), apoptosis covalent modification (Geiss-Friedlander and Melchior, 2007; is mainly mediated by endogenous and exogenous apoptotic Flotho and Melchior, 2013; Sheng et al., 2019). Three major pathways (Li, 1997; Li et al., 2001; Mao et al., 2001, 2004; Yao SUMO isoforms have been identified in vertebrates, which are et al., 2003; Kim and Koh, 2011). Bcl-2 family proteins play a known as SUMO1, SUMO2, and SUMO3 (Hay, 2005). crucial role in mediating endogenous apoptotic pathway, and Sumoylation is an enzymatic reaction catalyzed by the E1 among which Bax is one of the most important pro-apoptotic activating enzyme, the E2 conjugating enzyme, and several proteins (Mao et al., 2004; Czabotar et al., 2014; Singh et al., 2019; E3 ligases in the presence of ATP as energy supply, and Walensky, 2019; Moldoveanu and Czabotar, 2020). reversed by several SUMO isopeptidases known as SENPs In this study, we present evidence to show that during (Johnson, 2004; Hickey et al., 2012). Previous studies have oxidative stress-induced apoptosis, PIAS1 expression is shown that protein sumoylation plays an important role in significantly changed at the mRNA and protein levels. The regulating different biological processes including cell division, change in PIAS1 expression is closely related with apoptosis. autophagy, transformation, aging, and apoptosis (Johnson, PIAS1 knockout via CRISPR/Cas9 technology attenuates 2004; Hendriks and Vertegaal, 2016). Moreover, sumoylation oxidative stress-induced apoptosis. In contrast, expression of is also implicated in various human diseases caused by gene exogenous PIAS1 enhances oxidative stress-induced apoptosis mutation and stress response (Flotho and Melchior, 2013; of LECs. To define the underlying mechanism, we analyzed Seeler and Dejean, 2017; Yang et al., 2017; Sheng et al., 2019; the apoptosis-related factors in PIAS1 overexpression and Princz and Tavernarakis, 2020). knockout cells. Our results reveal that PIAS1 affects the During sumoylation reaction, E3 ligases have been reported expression level of the pro-apoptotic protein Bax. Furthermore, to stabilize the binding between the SUMO target proteins and we demonstrate that PIAS1 regulation of Bax occurs through the E2 conjugating enzyme, controlling the transfer of SUMO regulation of p53 sumoylation at the K386 residue in LECs. from E2 to substrate proteins (Hay, 2005; Hickey et al., 2012; The sumoylation-deficient p53 K386R mutant can protect cells Flotho and Melchior, 2013). Since the E1 activating enzyme against the stress-induced apoptosis. Finally, in Bax knockout (consisting of SAE1-UBA2 heterodimer) and the E2 conjugating cells, we found that absence of Bax significantly abrogates PIAS1- enzyme (UBC9) are the unique enzymes, the E3 ligases play a mediated apoptosis under oxidative stress induction. Taken key role in the selection of SUMO isoforms and target proteins together, our results demonstrate that PIAS1 is implicated in lens (Hay, 2005; Geiss-Friedlander and Melchior, 2007; Hendriks cataractogenesis. Mechanistically, PIAS1 sumoylates p53 at K386 and Vertegaal, 2016; Pichler et al., 2017). In mammals, the to upregulate Bax and thus promotes oxidative stress-induced protein inhibitor of activated STAT (PIAS) family of E3 ligases apoptosis through p53-Bax pathway. contain five subtypes, namely, PIAS1, PIAS2a, PIAS2b, PIAS3, and PIAS4, among which PIAS1 is known to promote cell apoptosis depending on its SUMO E3 ligase activity (Shuai and RESULTS Liu, 2005; Palvimo, 2007; Rytinki et al., 2009; Sudharsan and Azuma, 2012; Yang et al., 2013; Rabellino et al., 2017; Li et al., Oxidative Stress Induces PIAS1 2020). PIAS1 can regulate apoptosis by mediating sumoylation Alteration in Lens Epithelial Cells of both transcription factors and kinases (Liu and Shuai, 2001; It is well established that oxidative stress plays a causing role in Megidish et al., 2002; Shishido et al., 2008; Alm-Kristiansen cataractogenesis (Giblin et al., 1995; Li et al., 1995a,b; Spector, et al., 2011; Leitao et al., 2011; Sudharsan and Azuma, 2012; Li 1995; Li and Spector, 1996; Reddy et al., 2001; Raghavan et al., et al., 2013; Yang et al., 2013; Chiou et al., 2014; Wang et al., 2016; Rakete and Nagaraj, 2016; Barnes and Quinlan, 2017; Fan 2018) and thus becomes involved in various human diseases et al., 2017; Wang et al., 2017). In our glucose oxidase (GO) including cardiovascular diseases, neuronal disorder, and cancer treatment-induced cataract model (Supplementary Figure 1), we development (Fatkin et al., 1999; Sternsdorf et al., 1999; Li et al., found that GO also regulates PIAS1 expression. As shown in 2003; Steffan et al., 2004; Kim et al., 2006, 2007; Ballatore et al., Figure 1A, 40 mU GO induced time-dependent upregulation of 2007; Evdokimov et al., 2008; Subramaniam et al., 2009; Flotho PIAS1 mRNA in the first
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