Transcriptional modifications and the cytoprotective, DNA protective, and wound healing effects of peroxiredoxin-1 from Sebastes schlegelii

Rajamanthrilage Kasun Madusanka,M.D. Neranjan Tharuka,W.S.P. Madhuranga,Seongdo Lee,Jehee Lee 제주대학교 해양과학연구소 2020 해양과환경연구소 연구논문집 Vol.44 No.-

Peroxiredoxins are a group of thiol-specific antioxidant proteins that take six isoforms in vertebrates and allow the innate immune system to sense and detoxify reactive oxygen species. In this study, we identified and characterized the perxiredoxin-1 (SsPrdx1) cDNA sequence from the rockfish, Sebastes schlegelii. In silico analysis revealed that SsPrdx1 contained a 594 bp long open reading frame (ORF) encoding a protein of 198 amino acids, with a predicted molecular weight and theoretical isoelectric point of 21.97 kDa and 6.30, respectively. The SsPrdx1 gene comprised six exons linked by five introns, while peroxiredoxin signature motifs were found in the highly conserved third, fourth, and fifth exons. Phylogenetic analysis and sequence alignment suggested that SsPrdx1 is evolutionarily conserved and that its most closely related counterpart is Salarias fasciatus. Recombinant SsPrdx1 (rSsPrdx1) displayed supercoiled DNA protection and insulin disulfide reduction activities in a concentration-dependent manner, while cells transiently transfected with pcDNA3.1 (+)/SsPrdx1 exhibited significant cytoprotective effects under oxidative stress and wound healing activity. SsPrdx1 transcripts were constitutively expressed under normal physiological conditions, with the highest expression observed in the blood. Moreover, SsPrdx1 expression increased in the blood, spleen, and liver following immune provocation by LPS, poly I:C, and Streptococcus iniae injection. Thus, this study provides insights into the role of SsPrdx1 in rockfish immune protection.

상피성 난소암 세포주에서 1-Cys Peroxiredoxin의 발현과 Cisplatin에 대한 저항성과의 관계

주원덕 ( Won Duk Joo ),박장호 ( Jang Ho Pak ),김종혁 ( Jong Hyeok Kim ),이신형 ( Shin Hyung Lee ),김대연 ( Dae Yeon Kim ),서대식 ( Dae Shik Suh ),김용만 ( Yong Man Kim ),김영탁 ( Young Tak Kim ),목정은 ( Jung Eun Mok ),남주현 ( Jo 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.10

Objective: To investigate the relationship between expression of 1-Cys peroxiredoxin (Prx) and resistance to cisplatin in epithelial ovarian cancer cell lines. Methods: Immunohistochemistry of 1-Cys Prx was performed on both normal ovarian tissues and the tissues of epithelial ovarian cancer. Western blot was performed to measure the expression of 1-Cys Prx in SKOV-3, OVCAR-3 and SNU-8 after treatment with cisplatin. Expression of 1-Cys Prx in SKOV-3 was also measured according to both time after treatment with cisplatin and concentration of cisplatin. The generation of reactive oxygen species (ROS) was measured with and without antioxidants in SKOV-3. SKOV-3 was transfected with 1-Cys Prx green fluorescent protein plasmid to overexpress 1-Cys Prx and TUNEL assay was performed after treatment with cisplatin to examine apoptosis. Results: 1-Cys Prx was strongly expressed in both stroma and epithelium of both normal ovary and epithelial ovarian cancer, especially in the cytoplasm of epithelial cells. SNU-8 and OVCAR-3 exhibited about 1.5 fold higher expression than SKOV-3. SKOV-3 showed the peak expression at 48 hours after treatment with cisplatin and in 3 ㎍/㎖ concentration of cisplatin. The generation of ROS was increased after treatment with cisplatin to SKOV-3 and the survival of SKOV-3 against cisplatin was correlated with the concentration of antioxidants (p<0.001). No apoptosis occurred in 1-Cys Prx overexpressed SKOV-3 cells. Conclusion: 1-Cys Prx was shown to increase the resistance to cisplatin in epithelial ovarian cancer cell line. The result suggests that the resistance may be due to overexpression of 1-Cys Prx, which is responsible for removal of ROS generated by cisplatin.

Prdx1 (peroxiredoxin 1) deficiency reduces cholesterol efflux via impaired macrophage lipophagic flux

Jeong, Se-Jin,Kim, Sinai,Park, Jong-Gil,Jung, In-hyuk,Lee, Mi-Ni,Jeon, Sejin,Kweon, Hyae Yon,Yu, Dae-Yeul,Lee, Sang-Hak,Jang, Yangsoo,Kang, Sang Won,Han, Ki-Hwan,Miller, Yury I.,Park, Young Mi,Cheong, LANDES BIOSCIENCE 2018 AUTOPHAGY Vol.14 No.1

<P><B>ABSTRACT</B></P><P>Oxidative stress activates macroautophagy/autophagy and contributes to atherogenesis via lipophagic flux, a form of lipid removal by autophagy. However, it is not known exactly how endogenous antioxidant enzymes are involved in lipophagic flux. Here, we demonstrate that the antioxidant PRDX1 (peroxiredoxin 1) has a crucial role in the maintenance of lipophagic flux in macrophages. PRDX1 is more highly expressed than other antioxidant enzymes in monocytes and macrophages. We determined that <I>Prdx1</I> deficiency induced excessive oxidative stress and impaired maintenance of autophagic flux in macrophages. <I>Prdx1</I>-deficient macrophages had higher intracellular cholesterol mass and lower cholesterol efflux compared with wild type. This perturbation in cholesterol homeostasis was due to impaired lipophagic cholesterol hydrolysis caused by excessive oxidative stress, resulting in the inhibition of free cholesterol formation and the reduction of NR1H3 (nuclear receptor subfamily 1, group H, member 3) activity. Notably, impairment of both lipophagic flux and cholesterol efflux was restored by the 2-Cys PRDX-mimics ebselen and gliotoxin. Consistent with this observation, <I>apoe <SUP>−/−</SUP></I> mice transplanted with bone marrow from <I>prdx1<SUP>−/−</SUP>apoe<SUP>−/−</SUP></I> mice had increased plaque formation compared with <I>apoe<SUP>−/−</SUP></I> BM-transplanted recipients. This study reveals that PRDX1 is crucial to regulating lipophagic flux and maintaining macrophage cholesterol homeostasis against oxidative stress. We suggest that PRDX1-dependent control of oxidative stress may provide a strategy for treating atherosclerosis and autophagy-related human diseases.</P>

Transcriptional regulation of the murine 1-cys peroxiredoxin gene by the B cell-specific activator protein, Pax5

Lee, In-Seon,Choi, Won Ho,Kim, Ji Young,Jeong, Jin-Yong,Kim, Mi-Jung,Nam, Joo-Hyun,Kim, Jong-Hyeok,Seo, Sang-Beom,Pak, Jhang Ho Wiley Subscription Services, Inc., A Wiley Company 2008 Journal of cellular biochemistry Vol.104 No.2

<P>Pax5, a member of the paired box gene family of transcription factors, is a B cell-specific activator protein (BSAP) that plays important roles in controlling the expression of lineage- and differentiation-stage specific genes during B lymphopoiesis. We identified two putative Pax5 binding sites in a 668 bp of the murine 1-cys peroxiredoxin (1-cysPrx) promoter region. These sites were located at positions −278 to −262 and −50 to −34 from the translation start site. Gel mobility shift assays showed that recombinant Pax5 protein bound specifically to the nucleotide regions −56 to −24 (MP1 probe) and −284 to −253 (MP2 probe). Furthermore, endogenous Pax5 protein from B lymphoblast cells (IM-9) formed a DNA-protein complex with MP1 and MP2 probes, indicating that Pax5 binding occurs specifically at these sequences in vivo. Transient transfection studies showed that expression of exogenous Pax5 resulted in dose-dependent increases in 1-cysPrx promoter activity, implying that Pax5 functions as a positive transcription factor for 1-cysPrx expression. Further transient cotransfection studies showed that coexpression of Pax5 and histone acetyltransferases (HATs), such as p300, cAMP-response-element-binding protein (CBP) and p300/CBP associated factor (PCAF) enhanced the Pax5-mediated 1-cysPrx promoter activity. Immunoprecipitation studies indicated that Pax5 directly binds to HATs. Chromatin immunoprecipitation assays showed that the recruitment of Pax5 to the promoter induced histone H3 and H4 hyperacetylation of chromatin. Lipopolysaccharides (LPS) stimulation of murine splenocytes resulted in coordinated expression of Pax5 and 1-cysPrx proteins. These findings suggest that Pax5 may function as a transactivator of 1-cysPrx gene expression. J. Cell. Biochem. 104: 465–476, 2008. © 2008 Wiley-Liss, Inc.</P>

Effect of PRX-1 Downregulation in the Type 1 Diabetes Microenvironment

Yoo, Jong-Sun,Lee, Yun-Jung,Hyung, Kyeong Eun,Yoon, Joo Won,Lee, Ik Hee,Park, So-Young,Hwang, Kwang Woo The Korean Society of Pharmacology 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.6

Type 1 diabetes (T1D) is caused by dysregulation of the immune system in the pancreatic islets, which eventually leads to insulin-producing pancreatic ${\beta}$-cell death and destabilization of glucose homeostasis. One of the major characteristics of T1D pathogenesis is the production of inflammatory mediators by macrophages that result in destruction or damage of pancreatic ${\beta}$-cells. In this study the inflammatory microenvironment of T1D was simulated with RAW264.7 cells and MIN6 cells, acting as macrophages and pancreatic ${\beta}$-cells respectably. In this setting, peroxiredoxin-1, an anti-oxidant enzyme was knocked down to observe its functions in the pathogenesis of T1D. RAW264.7 cells were primed with lipopolysaccharide and co-cultured with MIN6 cells while PRX-1 was knocked down in one or both cell types. Our results suggest that hindrance of PRX-1 activity or the deficiency of this enzyme in inflammatory conditions negatively affects pancreatic ${\beta}$-cell survival. The observed decrease in viability of MIN6 cells seems to be caused by nitric oxide production. Additionally, it seems that PRX-1 affects previously reported protective activity of IL-6 in pancreatic ${\beta}$ cells as well. These results signify new, undiscovered roles for PRX-1 in inflammatory conditions and may contribute toward our understanding of autoimmunity.

Effect of PRX-1 Downregulation in the Type 1 Diabetes Microenvironment

Jong-Sun Yoo,이윤정,형경은,Joo Won Yoon,Ik Hee Lee,박소영,황광우 대한약리학회 2012 The Korean Journal of Physiology & Pharmacology Vol.16 No.6

Type 1 diabetes (T1D) is caused by dysregulation of the immune system in the pancreatic islets, which eventually leads to insulin-producing pancreatic β-cell death and destabilization of glucose homeostasis. One of the major characteristics of T1D pathogenesis is the production of inflammatory mediators by macrophages that result in destruction or damage of pancreatic β-cells. In this study the inflammatory microenvironment of T1D was simulated with RAW264.7 cells and MIN6 cells, acting as macrophages and pancreatic β-cells respectably. In this setting, peroxiredoxin-1, an anti-oxidant enzyme was knocked down to observe its functions in the pathogenesis of T1D. RAW264.7 cells were primed with lipopolysaccharide and co-cultured with MIN6 cells while PRX-1 was knocked down in one or both cell types. Our results suggest that hindrance of PRX-1 activity or the deficiency of this enzyme in inflammatory conditions negatively affects pancreatic β-cell survival. The observed decrease in viability of MIN6 cells seems to be caused by nitric oxide production. Additionally, it seems that PRX-1 affects previously reported protective activity of IL-6 in pancreatic β cells as well. These results signify new, undiscovered roles for PRX-1 in inflammatory conditions and may contribute toward our understanding of autoimmunity.

각질형성세포에서 UVB에 의한 Peroxiredoxin 1의 발현 조절: H2O2에 의한 Peroxiredoxin 1의 조절

윤숙정 ( Sook Jung Yun ),이재욱 ( Jae Wook Lee ),최지영 ( Jee Young Choi ),이승철 ( Seung Chul Lee ) 대한피부과학회 2005 大韓皮膚科學會誌 Vol.43 No.9

Over-expressed Peroxiredoxin 1 Protects against Oxidative Damage in Mouse Embryonic Fibroblasts Lacking Peroxiredoxin 2

( Sang Gyu Lee ),( Seong Gon Kim ),( Jae Young Kim ),( Zae Young Ryoo ) 한국응용약물학회 2011 Biomolecules & Therapeutics(구 응용약물학회지) Vol.19 No.4

Peroxiredoxins (Prxs) have a critical role in protecting cells against oxidative damage generated by reactive oxygen species (ROS). PrxI and PrxII are more than 90% homologous in their amino acid sequences, and both proteins reduce H2O2. In this study, an over-expression plasmid carrying PrxI was transfected into PrxII-/- mouse embryonic fi broblasts (MEFs) to investigate potential compensatory relationships between PrxI and PrxII. ROS levels induced by oxidative stress were increased in PrxII-/- MEFs as compared to wild-type MEFs. Moreover, exposure of PrxII-/- MEFs to H2O2 caused a reduction in cell viability of about 10%, and the proportion of cell death was increased compared to mock-treated PrxII-/- MEFs. However, transient over-expression of PrxI in PrxII-/- MEFs conferred increased resistance against the oxidative damage, as evidenced by increased cell viability and reduced intracellular ROS levels under H2O2 stress conditions. The fi ndings suggest that over-expressed PrxI can partly compensate for the loss of PrxII function in PrxII-defi cient MEFs.

Transcriptional Regulation of the AP-1 and Nrf2 Target Gene Sulfiredoxin

Francesc X. Soriano,Paul Baxter,Lyndsay M. Murray,Michael B. Sporn,Thomas H. Gillingwater,Giles E. Hardingham 한국분자세포생물학회 2009 Molecules and cells Vol.27 No.3

“Two-cysteine” peroxiredoxins are antioxidant enzymes that exert a cytoprotective effect in many models of oxidative stress. However, under highly oxidizing conditions they can be inactivated through hyperoxidation of their peroxidatic active site cysteine residue. Sulfiredoxin can reverse this hyperoxidation, thus reactivating peroxiredoxins. Here we review recent investigations that have shed further light on sulfiredoxin’s role and regulation. Studies have revealed sulfiredoxin to be a dynamically regulated gene whose transcription is induced by a variety of signals and stimuli. Sulfiredoxin expression is regulated by the transcription factor AP-1, which mediates its up-regulation by synaptic activity in neurons, resulting in protection against oxidative stress. Furthermore, sulfiredoxin has been identified as a new member of the family of genes regulated by Nuclear factor erythroid 2-related factor (Nrf2) via a conserved cis-acting antioxidant response element (ARE). As such, sulfiredoxin is likely to contribute to the net antioxidative effect of small molecule activators of Nrf2. As discussed here, the proximal AP-1 site of the sulfiredoxin promoter is embedded within the ARE, as is common with Nrf2 target genes. Other recent studies have shown that sulfiredoxin induction via Nrf2 may form an important part of the protective response to oxidative stress in the lung, preventing peroxiredoxin hyperoxidation and, in certain cases, subsequent degradation. We illustrate here that sulfiredoxin can be rapidly induced in vivo by administration of CDDO-TFEA, a synthetic triterpenoid inducer of endogenous Nrf2, which may offer a way of reversing peroxiredoxin hyperoxidation in vivo following chronic or acute oxidative stress.

백반증에서 표피내 Peroxiredoxin 1의 발현

전근 ( Gen Quan ),박미선 ( Mei Shan Piao ),최지영 ( Jee Young Choi ),전지선 ( Ji Sun Chun ),이지범 ( Jee Bum Lee ),이승철 ( Seung Chul Lee ) 대한피부과학회 2008 大韓皮膚科學會誌 Vol.46 No.6

Background: Although the pathogenesis of vitiligo isn`t fully understood, a recent study demonstrates that oxidative stress plays an important role to induce vitiligo. Peroxiredoxin (Prx) is a novel peroxidase family to remove hydrogen peroxide using thioredoxin system, which is consisted of thioredoxin, thioredoxin reductase, and NADPH. Objective: This study aimed to investigate the change of expression of Prx I to elucidate the role of oxidative stress in the pathogenesis of vitiligo. Methods: Sample specimens were obtained from the lesional skin of vitiligo patients, and non-depigmented skin was obtained from the perilesional area as control samples. The skin samples were immediately frozen using liquid nitrogen, and then section samples were prepared to perform immunohistochemical staining with antibodies for Prx I. Some of the skin biopsy samples were used for primary culture of keratinocytes. Protein extracts from the expanded keratinocytes were prepared for Western blot analysis of Prx I. Results: In vitiligo, the ubiquitous expression of Prx I in all layers of epidermis, which was also observed in the normal perilesional skin, was reduced in the depigmented lesion of vitiligo patients. The reduction of Prx I was remarkable from the lesions which were exposed to sunlight. Consistently, Prx I expression from the lesional keratinocytes were noticeably reduced in comparison with that from perilesional keratinocytes. Conclusion: Our results showing that Prx I is impaired in the epidermis of depigmented lesions of vitiligo patients suggest that oxidative stress is an important factor to induce vitiligo. (Korean J Dermatol 2008;46(6):736∼741)