| 55 | 0 | 96 |
| 下载次数 | 被引频次 | 阅读次数 |
激活转录因子4(Activating transcription factor 4, ATF4)是参与机体应激调控的重要转录因子,对细胞代谢具有关键调控作用。斑马鱼基因组包含atf4a和atf4b两个亚型,目前已有关于Atf4a功能的研究报道,而Atf4b的功能尚未明确。通过序列比对,本文发现斑马鱼Atf4b的序列仅在亮氨酸拉链结构域和DNA结合结构域与哺乳动物ATF4序列保持相对一致。为探究Atf4b对斑马鱼生长发育及代谢功能的影响,作者通过CRISPR/Cas9技术构建斑马鱼atf4b基因敲除品系。形态学研究表明,atf4b基因敲除(atf4b-/-)胚胎无明显畸形,但显著减少斑马鱼胚胎体长。代谢组学分析发现,突变体的三羧酸循环中柠檬酸含量升高,而下游代谢物(α-酮戊二酸、苹果酸、琥珀酸)减少,ATP生成显著降低。Atf4b的缺失造成线粒体损伤,并且显著降低酸胁迫下的存活率。本研究证实Atf4b在斑马鱼生长发育及能量代谢过程中发挥重要作用,研究结果为深入研究斑马鱼的发育、代谢和应激调控耦合机制提供了新视角。
Abstract:Activating transcription factor 4(ATF4) is a critical transcription factor involved in stress response. It exerts essential regulatory effects on cellular metabolism. Zebrafish genome contains two ATF4 paralogs, atf4a and atf4b. The function of Atf4a has been documented previously while that of Atf4b remains elusive. To investigate the impact of Atf4b on zebrafish development and metabolism, we generated an atf4b knockout zebrafish using CRISPR/Cas9. Sequence alignment revealed that zebrafish Atf4b shares conserved domains only in the leucine zipper and DNA-binding regions with mammalian ATF4. Morphological examination showed no apparent deformities in atf4b-/- embryos while body length was significantly reduced. Metabolomic profiling demonstrated the elevated citrate level accompanied by reduced downstream metabolites like oxoglutarate, malate and succinate in the tricarboxylic acid(TCA) cycle with a marked decrease in ATP production. Atf4b deficiency induced mitochondrial damage and significantly compromised survival under acidic stress conditions. These findings evidenced the crucial role of Atf4b in zebrafish growth, energy metabolism and stress adaptation, and provided novel insights into the integrated mechanism coupling developmental processes, metabolic homeostasis and stress response.
[1] Lu H J,Koju N,Sheng R.Mammalian integrated stress responses in stressed organelles and their functions[J].Acta Pharmacologica Sinica,2024,45:1095-1114.
[2] Ameri K,Harris A L.Activating transcription factor 4[J].The International Journal of Biochemistry & Cell Biology,2008,40:14-21.
[3] hang N,Zhang S,Dong X.Plant-derived bioactive compounds and their novel role in central nervous system disorder treatment via ATF4 targeting:A systematic literature review[J].Biomed Pharmacother,2024,176:116811.
[4] Xiao Y,Xie X,Chen Z,et al.Advances in the roles of ATF4 in osteoporosis[J].Biomed Pharmacother,2023,169:115864.
[5] Li M,Tang S,Velkov T,et al.Copper exposure induces mitochondrial dysfunction and hepatotoxicity via the induction of oxidative stress and PERK/ATF4 -mediated endoplasmic reticulum stress[J].Environmental Pollution,2024,352:124145.
[6] Takahashi M,Okamoto Y,Kato Y,et al.Activating mutations in EGFR and PI3K promote ATF4 induction for NSCLC cell survival during amino acid deprivation[J].Heliyon,2023,9:e14799.
[7] Kilberg M S,Shan J,Su N.ATF4-dependent transcription mediates signaling of amino acid limitation[J].Trends in Endocrinology & Metabolism,2009,20:436-443.
[8] B′Chir W,Maurin A C,Carraro V,et al.The eIF2α/ATF4 pathway is essential for stress-induced autophagy gene expression[J].Nucleic Acids Research,2013,41:7683-7699.
[9] Harding H P,Novoa I,Zhang Y,et al.Regulated translation initiation controls stress-induced gene expression in mammalian cells[J].Molecular Cell,2000,6:1099-1108.
[10] Calfon M,Zeng H,Urano F,et al.IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA[J].Nature,2002,415:92-96.
[11] Jousse C,Deval C,Maurin A C,et al.TRB3 inhibits the transcriptional activation of stress-regulated genes by a negative feedback on the ATF4 pathway[J].Journal of Biological Chemistry,2007,282:15851-15861.
[12] Ye J,Kumanova M,Hart L S,et al.The GCN2-ATF4 pathway is critical for tumour cell survival and proliferation in response to nutrient deprivation[J].The EMBO Journal,2010,29:2082-2096.
[13] Long F.Building strong bones:Molecular regulation of the osteoblast lineage[J].Nature Reviews Molecular Cell Biology,2011,13:27-38.
[14] Salaroglio I C,Panada E,Moiso E,et al.PERK induces resistance to cell death elicited by endoplasmic reticulum stress and chemotherapy[J].Molecular Cancer,2017,16:91.
[15] Jeon W J,Kim K M,Kim E J,et al.Costunolide increases osteoblast differentiation via ATF4-dependent HO-1 expression in C3H10T1/2 cells[J].Life Sciences,2017,178:94-99.
[16] Chen D,Fan Z,Rauh M,et al.ATF4 promotes angiogenesis and neuronal cell death and confers ferroptosis in a xCT-dependent manner[J].Oncogene,2017,36:5593-5608.
[17] Cui A,Ding D,Li Y.Regulation of hepatic metabolism and cell growth by the ATF/CREB family of transcription factors[J].Diabetes,2021,70:653-664.
[18] Chen C,Zhang Z,Liu C,et al.ATF4-dependent fructolysis fuels growth of glioblastoma multiforme[J].Nature Communications,2022,13:6108.
[19] Heinemann-Yerushalmi L,Bentovim L,Felsenthal N,et al.BCKDK regulates the TCA cycle through PDC in the absence of PDK family during embryonic development[J].Developmental Cell,2021,56:1182-1194.
[20] Kang W,Suzuki M,Saito T,et al.Emerging role of TCA cycle-related enzymes in human diseases[J].International Journal of Molecular Sciences,2021,22:313057.
[21] Martínez-Reyes I,Chandel N S.Mitochondrial TCA cycle metabolites control physiology and disease[J].Nature Communications,2020,11:102.
[22] Martínez-Reyes I,Diebold L P,Kong H,et al.TCA cycle and mitochondrial membrane potential are necessary for diverse biological functions[J].Molecular Cell,2016,61:199-209.
[23] Brière J J,Favier J,Gimenez-Roqueplo A P,et al.Tricarboxylic acid cycle dysfunction as a cause of human diseases and tumor formation[J].American Journal of Physiology,2006,291:1114-1120.
[24] Fusakio M E,Willy J A,Wang Y,et al.Transcription factor ATF4 directs basal and stress-induced gene expression in the unfolded protein response and cholesterol metabolism in the liver[J].Molecular Biology of the Cell,2016,27:1536-1551.
[25] Wang C,Huang Z,Du Y,et al.ATF4 regulates lipid metabolism and thermogenesis[J].Cell Research,2010,20:174-184.
[26] Li H,Meng Q,Xiao F,et al.ATF4 deficiency protects mice from high-carbohydrate-diet-induced liver steatosis[J].Biochemical Journal,2011,438:283-289.
[27] Masuoka H C,Townes T M.Targeted disruption of the activating transcription factor 4 gene results in severe fetal anemia in mice[J].Blood,2002,99:736-745.
[28] Yeh K Y,Lai C Y,Lin C Y,et al.ATF4 overexpression induces early onset of hyperlipidaemia and hepatic steatosis and enhances adipogenesis in zebrafish[J].Scientific Reports,2017,7:16362.
[29] Petel Légaré V,Rampal C J,Gurberg T J N,et al.Loss of mitochondrial Chchd10 or Chchd2 in zebrafish leads to an ALS-like phenotype and complex Ⅰ deficiency independent of the mitochondrial integrated stress response[J].Developmental Neurobiology,2023,83:54-69.
[30] Doll L,Welte K,Skokowa J,et al.A JAGN1-associated severe congenital neutropenia zebrafish model revealed an altered G-CSFR signaling and UPR activation[J].Blood Advances,2024,8:4050-4065.
[31] Hwang W Y,Fu Y,Reyon D,et al.Efficient genome editing in zebrafish using a CRISPR-Cas system[J].Nature Biotechnology,2013,31:227-229.
[32] Tuladhar R,Yeu Y,Tyler Piazza J,et al.CRISPR-Cas9-based mutagenesis frequently provokes on-target mRNA misregulation[J].Nature Communications,2019,10:4056.
[33] Zhang Q,Yu J,Liu B,et al.Central activating transcription factor 4 (ATF4) regulates hepatic insulin resistance in mice via S6K1 signaling and the vagus nerve[J].Diabetes,2013,62:2230-2239.
[34] Hiwatashi Y,Kanno K,Takasaki C,et al.PHD1 interacts with ATF4 and negatively regulates its transcriptional activity without prolyl hydroxylation[J].Experimental Cell Research,2011,317:2789-2799.
[35] Neill G,Masson G R.A stay of execution:ATF4 regulation and potential outcomes for the integrated stress response[J].Frontiers in Molecular Neuroscience,2023,16:1112253.
[36] Torrence M E,MacArthur M R,Hosios A M,et al.The mTORC1-mediated activation of ATF4 promotes protein and glutathione synthesis downstream of growth signals[J].Elife,2021,10:e63326.
[37] Castellani C A,Longchamps R J,Sun J,et al.Thinking outside the nucleus:Mitochondrial DNA copy number in health and disease[J].Mitochondrion,2020,53:214-223.
[38] Guha M,Avadhani N G.Mitochondrial retrograde signaling at the crossroads of tumor bioenergetics,genetics and epigenetics[J].Mitochondrion,2013,13:577-591.
[39] Abd Radzak S M,Mohd Khair S Z N,Ahmad F,et al.Insights regarding mitochondrial DNA copy number alterations in human cancer (Review)[J].International Journal of Molecular Medicine,2022,50:104.
[40] Hu L,Yao X,Shen Y.Altered mitochondrial DNA copy number contributes to human cancer risk:evidence from an updated meta-analysis[J].Scientific Reports,2016,6:35859.
[41] Lee H,Lee J H,Kim D C,et al.Is mitochondrial DNA copy number associated with clinical characteristics and prognosis in gastric cancer?[J].Asian Pacific Journal of Cancer Prevention,2015,16:87-90.
[42] Palmieri F,Pierri C L.Mitochondrial metabolite transport[J].Essays in Biochemistry,2010,47:37-52.
[43] Ternette N,Yang M,Laroyia M,et al.Inhibition of mitochondrial aconitase by succination in fumarate hydratase deficiency[J].Cell Reports,2013,3:689-700.
[44] Lushchak O V,Piroddi M,Galli F,et al.Aconitase post-translational modification as a key in linkage between Krebs cycle,iron homeostasis,redox signaling,and metabolism of reactive oxygen species[J].Redox Report,2014,19:8-15.
[45] Fernandez H R,Gadre S M,Tan M,et al.The mitochondrial citrate carrier,SLC25A1,drives stemness and therapy resistance in non-small cell lung cancer[J].Cell Death & Differentiation,2018,25:1239-1258.
[46] Li Z,Wang H.Molecular mechanisms of the SLC13A5 gene transcription[J].Metabolites,2021,11:706.
[47] Hallan S,Afkarian M,Zelnick L R,et al.Metabolomics and gene expression analysis reveal down-regulation of the citric acid (TCA) cycle in non-diabetic CKD patients[J].eBioMedicine,2017,26:68-77.
[48] Volz K.The functional duality of iron regulatory protein 1[J].Current Opinion in Structural Biology,2008,18:106-111.
[49] He Q,Yu T,Chen J,et al.Enhancement of de novo lipogenesis by the IDH1 and IDH2-dependent reverse TCA cycle maintains the growth and angiogenic capacity of bone marrow-derived endothelial progenitor cells under hypoxia[J].Free Radical Biology and Medicine,2024,213:327-342.
[50] Feng X,Zhang L,Xu S,et al.ATP-citrate lyase (ACLY) in lipid metabolism and atherosclerosis:An updated review[J].Progress in Lipid Research,2020,77:101006.
[51] Verschueren K H G,Blanchet C,Felix J,et al.Structure of ATP citrate lyase and the origin of citrate synthase in the Krebs cycle[J].Nature,2019,568:571-575.
[52] Son S M,Park S J,Lee H,et al.Leucine signals to mTORC1 via its metabolite acetyl-coenzyme A[J].Cell Metabolism,2019,29:192-201.
[53] Lu S,Deng R,Jiang H,et al.The mechanism of ATP-Dependent allosteric protection of akt kinase phosphorylation[J].Structure,2015,23:1725-1734.
[54] Cai Z,Li C F,Han F,et al.Phosphorylation of PDHA by AMPK drives TCA cycle to promote cancer metastasis[J].Molecular Cell,2020,80:263-278.
[55] Cai Z,Peng D,Lin H K.AMPK maintains TCA cycle through sequential phosphorylation of PDHA to promote tumor metastasis[J].Cell Stress,2020,4:273-277.
[56] Tang X,Lucas J E,Chen J L,et al.Functional interaction between responses to lactic acidosis and hypoxia regulates genomic transcriptional outputs[J].Cancer Research,2012,72:491-502.
[57] Johno H,Ogata R,Nakajima S,et al.Acidic stress-ER stress axis for blunted activation of NF-κB in mesothelial cells exposed to peritoneal dialysis fluid[J].Nephrology Dialysis Transplantation,2012,27:4053-4060.
[58] Aoyama K,Burns D M,Suh S W,et al.Acidosis causes endoplasmic reticulum stress and caspase-12-mediated astrocyte death[J].Journal of Cerebral Blood Flow & Metabolism,2005,25:358-370.
[59] Williams C H,Neitzel L R,Cornell J,et al.GPR68-ATF4 signaling is a novel prosurvival pathway in glioblastoma activated by acidic extracellular microenvironment[J].Experimental Hemato-logy & Oncology,2024,13:13.
[60] McQuiston A,Diehl J A.Recent insights into PERK-dependent signaling from the stressed endoplasmic reticulum[J].F1000Research,2017,6:1897.
[61] Khacho M,Tarabay M,Patten D,et al.Acidosis overrides oxygen deprivation to maintain mitochondrial function and cell survival[J].Nature Communications,2014,5:3550.
[62] Kreß J K C,Jessen C,Hufnagel A,et al.The integrated stress response effector ATF4 is an obligatory metabolic activator of NRF2[J].Cell Reports,2023,42:112724.
基本信息:
DOI:10.16441/j.cnki.hdxb.20250133
中图分类号:S917.4
引用信息:
[1]夏伟意,王旋,周慧慧,等.Atf4b对斑马鱼三羧酸循环代谢和胚胎早期发育的调控作用[J].中国海洋大学学报(自然科学版),2026,56(07):43-52.DOI:10.16441/j.cnki.hdxb.20250133.
基金信息:
国家自然科学基金项目(32002365); 国家现代农业产业技术体系项目(CARS-47-G10)资助~~
2025-04-14
2025
2025-05-29
2025-12-26
2025
1
2026-06-22
2026-06-22