PMID- 36096331 OWN - NLM STAT- MEDLINE DCOM- 20221004 LR - 20221004 IS - 1879-0038 (Electronic) IS - 0378-1119 (Linking) VI - 847 DP - 2022 Dec 30 TI - MafG-like contribute to copper and cadmium induced antioxidant response by regulating antioxidant enzyme in Procambarus clarkii. PG - 146848 LID - S0378-1119(22)00667-9 [pii] LID - 10.1016/j.gene.2022.146848 [doi] AB - Avian musculoaponeurotic fibrosarcoma (Maf) proteins play an important role in Nrf2/Keap1 signaling pathway, which mainly resist the oxidant stress. The members of sMaf have a high homology basic leucine zipper (bZIP) and lack trans activation domain, and could interact with other transcriptional regulatory factors as a molecular chaperone. In this study, a full-length MafG-like gene was cloned from Procambarus Clarkii, designated as PcMafG-like, which consisted of an ORF length of 246 bp encoding 82 amino acids, a 5' untranslated region (UTR) of 483 bp, and a 3' UTR of 111 bp. The domain of PcMafG-like had a bZIP-Maf domain that binds to DNA. The cDNA sequence of PcMafG-like was 99 % similar to that of Penaeus vannamei. The mRNA of PcMafG-like was expressed in all tested tissues, and the highest expression was in muscle tissue. Under stimulation of Cu(2+) and Cd(2+), PcMafG-like was significantly up-regulated in hepatopancreas and gill, and the same result was testified by situ hybridization. The representative antioxidant genes, CAT, GPx and CZ-SOD, were significantly induced by Cu(2+); CAT and GPx was induced by Cd(2+). PcMafG-dsRNA significantly inhibited the expression of these up-regulated genes, but also inhibited the expression of other detected genes CZ-SOD, GST-theta and GST-1like. The antioxidant effect of PcMafG-like was further verified by oxidative stress markers (T-SOD, CuZnSOD, GPx, CAT, GSH and MDA) kits. Cu(2+) and Cd(2+) could induce the contents of these oxidative stress markers (MDA, GSH, CZ-SOD, CAT in Cu(2+)/Cd(2+) treated group, and GSH-Px in Cd(2+) group), while interference of PcMafG-like significantly inhibited the up-regulation. Furthermore, hematoxylin-eosin staining experiments showed that the degree of pathological damage was dose-dependent and time-dependent, and the pathological damage was more serious after dsRNA interfered with PcMafG-like. In addition, subcellular localization showed that PcMafG-like gene existed in nucleus. The recombinant protein PcMafG-like was expressed and purified in prokaryotic expression. The affinity analysis of promoter by agarose gel electrophoresis suggested that PcMafG-like could bind with CAT promoter in vitro. This indicated that PcMafG-like could activate antioxidant genes. CI - Copyright (c) 2022. Published by Elsevier B.V. FAU - Wang, Chengli AU - Wang C AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. FAU - Hu, Huogen AU - Hu H AD - Hydrobiology Protection and Rescue Center of Jiangxi, Nanchang 330029, China. FAU - Zeng, Liugen AU - Zeng L AD - Nanchang Academy of Agricultural Sciences, Nanchang 330038, China. FAU - Ni, Caiying AU - Ni C AD - College of Geography and Enviorment, Jiangxi Normal University, Nanchang 330033, China. FAU - An, Jinhua AU - An J AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. FAU - Gang, Yang AU - Gang Y AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. FAU - Jian, Shaoqing AU - Jian S AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. FAU - Wen, Chungen AU - Wen C AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. Electronic address: cgwen@ncu.edu.cn. FAU - Hu, Baoqing AU - Hu B AD - College of Life Science, Education Ministry Key Laboratory of Poyang Lake Environment and Resource Utilization, Nanchang University, Nanchang 330031, China. Electronic address: baoqinghu@sina.com. LA - eng PT - Journal Article DEP - 20220909 PL - Netherlands TA - Gene JT - Gene JID - 7706761 RN - 0 (3' Untranslated Regions) RN - 0 (5' Untranslated Regions) RN - 0 (Amino Acids) RN - 0 (Antioxidants) RN - 0 (DNA, Complementary) RN - 0 (Kelch-Like ECH-Associated Protein 1) RN - 0 (NF-E2-Related Factor 2) RN - 0 (Oxidants) RN - 0 (Recombinant Proteins) RN - 0 (Water Pollutants, Chemical) RN - 00BH33GNGH (Cadmium) RN - 789U1901C5 (Copper) RN - EC 1.15.1.1 (Superoxide Dismutase) RN - TDQ283MPCW (Eosine Yellowish-(YS)) RN - YKM8PY2Z55 (Hematoxylin) SB - IM MH - 3' Untranslated Regions MH - 5' Untranslated Regions MH - Amino Acids/genetics MH - Animals MH - *Antioxidants/pharmacology MH - Astacoidea/genetics MH - Cadmium/metabolism MH - Copper/pharmacology MH - DNA, Complementary/metabolism MH - Eosine Yellowish-(YS)/metabolism/pharmacology MH - Hematoxylin/metabolism/pharmacology MH - Kelch-Like ECH-Associated Protein 1/genetics MH - NF-E2-Related Factor 2/genetics MH - Oxidants/metabolism MH - Oxidative Stress MH - Recombinant Proteins/genetics MH - Superoxide Dismutase/genetics MH - *Water Pollutants, Chemical/metabolism OTO - NOTNLM OT - Antioxidant genes OT - CAT OT - Cadmium OT - Copper OT - MafG OT - Procambarus clarkii COIS- Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2022/09/13 06:00 MHDA- 2022/10/05 06:00 CRDT- 2022/09/12 19:26 PHST- 2022/03/28 00:00 [received] PHST- 2022/07/11 00:00 [revised] PHST- 2022/08/25 00:00 [accepted] PHST- 2022/09/13 06:00 [pubmed] PHST- 2022/10/05 06:00 [medline] PHST- 2022/09/12 19:26 [entrez] AID - S0378-1119(22)00667-9 [pii] AID - 10.1016/j.gene.2022.146848 [doi] PST - ppublish SO - Gene. 2022 Dec 30;847:146848. doi: 10.1016/j.gene.2022.146848. Epub 2022 Sep 9.