Molecular mechanisms of Mmd2 gene in regulating growth of the Pacific white shrimp Litopenaeus vannamei

Abstract

Growth of the Pacific white shrimp Litopenaeus vannamei, the most important farmed crustacean, has consistently been a focal point for breeders. Over the past decades, some candidate genes for shrimp growth have been identified. However, further research is needed to elucidate the molecular regulatory mechanism of these genes. LvMmd2 was previously identified as a candidate gene that may inhibit the growth of L. vannamei. In this study, we analyzed the genotype and expression of the LvMmd2 gene in a breeding family and indicated its role as a growth-inhibiting gene. We found that LvMmd2 co-localized with its homolog LvPAQR3 at the Golgi apparatus. Using co-immunoprecipitation (Co-IP) and DUAL membrane system yeast two-hybrid (MbY2H), we indicated the interactions between LvMmd2 and LvPAQR3, LvPAQR3 and LvRaf1, as well as LvMmd2 and LvRho. These results suggest that LvMmd2 directly and indirectly regulates the Ras signaling pathway. Furthermore, we show that the LvMmd2 gene may indirectly affect the PI3K/AKT, insulin, and Hippo signaling pathways to regulate cell proliferation and differentiation via LvPAQR3 and LvRaf1. Through transcriptome and MbY2H analyses, we have also revealed the interaction between LvMmd2 and proteins involved in growth, immunity, protein transport, synthesis, and modification. These findings demonstrate the various molecular pathways through which LvMmd2 regulates L. vannamei growth. This study provides insights into the mechanism of shrimp growth regulated by Mmd2, enhances our understanding of LvMmd2 function, and highlights its potential application in shrimp breeding.

Keywords

Pacific white shrimp / Monocyte to macrophage differentiation factor 2 / Growth-inhibiting gene / Function / Ras signaling pathway

References

[1] Aihara A, Iwawaki T, Abe-Fukasawa N, Otsuka K, Saruhashi K, Mikashima T, Nishino T. Small molecule LATS kinase inhibitors block the Hippo signaling pathway and promote cell growth under 3D culture conditions. J Biol Chem. 2022, 298 101779

[2] Alavi A, Acevedo L, Min W, Cheresh D. Chemoresistance of endothelial cells induced by basic fibroblast growth factor depends on Raf-1-mediated inhibition of the proapoptotic kinase, ASK1. Cancer Res. 2007, 67: 2766-2772

[3] Baida GE, Kuzmin NP. Cloning and primary structure of a new hemolysin gene from Bacillus cereus. Biochim Biophys Acta. 1995, 1264: 151-154

[4] Baida GE, Kuzmin NP. Mechanism of action of hemolysin III from Bacillus cereus. Biochim Biophys Acta. 1996, 1284: 122-124

[5] Bordbar F, Jensen J, Abied A, Guo W, Xu L, Gao H-J, Zhang L, Li J. Identification and validation of a novel candidate gene regulating net meat weight in Simmental beef cattle based on imputed next-generation sequencing. Cell Prolif. 2020, 53 e12870

[6] Cass LA, Meinkoth JL. Differential effects of cyclic adenosine 3',5'-monophosphate on p70 ribosomal S6 kinase. Endocrinology. 1998, 139: 1991-1998

[7] Chen J, Fujii K, Zhang L, Roberts T, Fu H. Raf-1 promotes cell survival by antagonizing apoptosis signal-regulating kinase 1 through a MEK-ERK independent mechanism. Proc Natl Acad Sci USA. 2001, 98: 7783-7788

[8] Cheng J, Chan TY, Zhang N, Sun S, Zhongli S. Mitochondrial phylogenomics reveals insights into taxonomy and evolution of penaeoidea (Crustacea: Decapoda). Zool Scr. 2018, 47: 582-594

[9] Crespo P, Leon J. Ras proteins in the control of the cell cycle and cell differentiation. Cell Mol Life Sci. 2000, 57: 1613-1636

[10] De Grave S, Fransen C. Carideorum catalogus: the recent species of the dendrobranchiate, stenopodidean, procarididean and caridean shrimps (Crustacea: Decapoda). Zoologische Mededelingen. 2011, 85: 195-588

[11] Dhillon AS, Meikle S, Yazici Z, Eulitz M, Kolch W. Regulation of Raf-1 activation and signalling by dephosphorylation. EMBO J. 2002, 21: 64-71

[12] Ehrenreiter K, Piazzolla D, Velamoor V, Sobczak I, Small J, Takeda J, Leung T, Baccarini M. Raf-1 regulates rho signaling and cell migration. J Cell Biol. 2005, 168: 955-964

[13] Ehrenreiter K, Kern F, Velamoor V, Meissl K, Galabova G, Sibilia M, Baccarini M. Raf-1 addiction in Ras-induced skin carcinogenesis. Cancer Cell. 2009, 16: 149-160

[14] Fan F, Feng L, He J, Wang X, Jiang X, Zhang Y, Wang Z, Chen Y. RKTG sequesters B-Raf to the golgi apparatus and inhibits the proliferation and tumorigenicity of human malignant melanoma cells. Carcinogenesis. 2008, 29: 1157-1163

[15] Feng L, Xie X, Ding Q, Luo X, He J, Fan F, Liu W, Wang Z, Chen Y. Spatial regulation of Raf kinase signaling by RKTG. Proc Natl Acad Sci USA. 2007, 104: 14348-14353

[16] García-Gutiérrez L, Fallahi E, Aboud N, Quinn N, Matallanas D. Interaction of LATS1 with SMAC links the MST2/Hippo pathway with apoptosis in an IAP-dependent manner. Cell Death Dis. 2022, 13: 692

[17] Hayashi T, Takeshita K, Tsuchida N, Kitano K, Kawabata S, Iwanaga S, Ito T. Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle. Int J Biochem Cell Biol. 1998, 30: 897-908

[18] Hayashi T, Ogawa T, Sato M, Tsuchida N, Fotovati A, Iwamoto H, Ikeuchi Y, Cassens RG, Ito T. S-myotrophin promotes the hypertrophy of myotube as insulin-like growth factor-I does. Int J Biochem Cell Biol. 2001, 33: 831-838

[19] Hindley A, Kolch W. Raf-1 and B-Raf promote protein kinase C theta interaction with BAD. Cell Signal. 2007, 19: 547-555

[20] Hüser M, Luckett J, Chiloeches A, Mercer K, Iwobi M, Giblett S, Sun XM, Brown J, Marais R, Pritchard C. MEK kinase activity is not necessary for Raf-1 function. EMBO J. 2001, 20: 1940-1951

[21] Jin T, Ding Q, Huang H, Xu D, Jiang Y, Zhou B, Li Z, Jiang X, He J, Liu W, Zhang Y, Pan Y, Wang Z, Thomas WG, Chen Y. PAQR10 and PAQR11 mediate Ras signaling in the Golgi apparatus. Cell Res. 2012, 22: 661-676

[22] Jin T, Xu D, Ding Q, Zhang Y, Mao C, Pan Y, Wang Z, Chen Y. Identification of the topology and functional domains of PAQR10. Biochem J. 2012, 443: 643-653

[23] Jin Y, Ha N, Forés M, Xiang J, Gläßer C, Maldera J, Jimenez G, Edgar B. EGFR/Ras signaling controls Drosophila intestinal stem cell proliferation via capicua-regulated genes. PLoS Genet. 2015, 11 e1005634

[24] Kamimura M, Meier K, Cavalli R, Laurino J, Maggioni R, Marins L. Characterization of growth-related genes in the south-western Atlantic pink shrimp Farfantepenaeus paulensis (Pérez-Farfante 1967) through a modified DDRT-PCR protocol. Aquac Res. 2008, 39: 200-204

[25] Khan AQ, Kuttikrishnan S, Siveen KS, Prabhu KS, Shanmugakonar M, Al-Naemi HA, Haris M, Dermime S, Uddin S. Ras-mediated oncogenic signaling pathways in human malignancies. Semin Cancer Biol. 2019, 54: 1-13

[26] Khosravi-Far R, Der CJ. The ras signal transduction pathway. Cancer Metastasis Rev. 1994, 13: 67-89

[27] Lee SJ, McPherron AC. Regulation of myostatin activity and muscle growth. Proc Natl Acad Sci USA. 2001, 98: 9306-9311

[28] Lee JY, Hopkinson NS, Kemp PR. Myostatin induces autophagy in skeletal muscle in vitro. Biochem Biophys Res Commun. 2011, 415: 632-636

[29] Li D-H, Chan T, Satow R, Komozaki S, Hashizume K, Asashima M. The role of XTRAP-gamma in Xenopus pronephros development. Int J Dev Biol. 2005, 49: 401-448

[30] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods. 2001, 25: 402-408

[31] Lopes-Ferreira M, Sosa-Rosales I, da Silva JP, Conceição K, Maleski A, Balan-Lima L, Disner G, Lima C. Molecular characterization and functional analysis of the nattectin-like toxin from the venomous fish Thalassophryne maculosa. Toxins (Basel). 2021, 14: 2

[32] Lyu D, Yu Y, Wang Q, Luo Z, Zhang Q, Zhang X, Xiang J, Li F. Identification of growth-associated genes by genome-wide association study and their potential application in the breeding of pacific white shrimp (Litopenaeus vannamei). Front Genet. 2021, 12 611570

[33] Nolan A, Aboud N, Kolch W, Matallanas D. Hidden targets in Raf signalling pathways to block oncogenic Ras signalling. Genes. 2021, 12: 553

[34] O'Neill E, Rushworth L, Baccarini M, Kolch W. Role of the kinase MST2 in suppression of apoptosis by the proto-oncogene product Raf-1. Science. 2005, 306: 2267-2270

[35] Pan P-L, Ye Y-X, Lou Y-H, Lu J-B, Cheng C, Shen Y, Moussian B, Zhang C-X. A comprehensive omics analysis and functional survey of cuticular proteins in the brown planthopper. Proc Natl Acad Sci USA. 2018, 115: 5175-5180

[36] Qiao S, Yang D, Li X, Li W, Zhang Y, Liu W. Silencing PAQR3 protects against oxygen-glucose deprivation/reperfusion-induced neuronal apoptosis via activation of PI3K/AKT signaling in PC12 cells. Life Sci. 2021, 265 118806

[37] Qin Y, Luo Z, Zhao K, Nan X, Guo Y, Li W, Wang Q. A new SVWC protein functions as a pattern recognition protein in antibacterial responses in Chinese mitten crab (Eriocheirsinensis). Fish Shellfish Immunol. 2022, 131: 1125-1135

[38] Shiraishi S, Nakamura Y-N, Iwamoto H, Haruno A, Sato Y, Mori S, Ikeuchi Y, Chikushi J, Hayashi T, Sato M, Cassens RG, Ito T. S-myotrophin promotes the hypertrophy of skeletal muscle of mice in vivo. Int J Biochem Cell Biol. 2006, 38: 1114-1122

[39] Si S, Zhang X, Yu Y, Zhang X, Zhong X, Yuan J, Yang S, Li F. Structure and function analyses of the Mmd2 gene in pacific white shrimp Litopenaeus vannamei. Front Genet. 2023, 14: 1151193

[40] Storch J, Thumser AE. Tissue-specific functions in the fatty acid-binding protein family. J Biol Chem. 2010, 285: 32679-32683

[41] Tang YT, Hu T, Arterburn M, Boyle B, Bright JM, Emtage PC, Funk WD. PAQR proteins: a novel membrane receptor family defined by an ancient 7-transmembrane pass motif. J Mol Evol. 2005, 61: 372-380

[42] Thomas P, Pang Y, Dong J, Groenen P, Kelder J, Vlieg J, Zhu Y, Tubbs C. Steroid and G protein binding characteristics of the seatrout and human progestin membrane receptor alpha subtypes and their evolutionary origins. Endocrinology. 2007, 148: 705-718

[43] Wang H-G, Rapp UR, Reed JC. Bcl-2 targets the protein kinase Raf-1 to mitochondria. Cell. 1996, 87: 629-638

[44] Wang L, Zhang J, Kong X, Zhao X, Pei C, Li L. A c-type lectin, nattectin-like protein (CaNTC) in qihe crucian carp Carassius auratus: binding ability with LPS, PGN and various bacteria, and agglutinating activity against bacteria. Fish Shellfish Immunol. 2017, 67: 382-392

[45] Wang Q, Yu Y, Zhang Q, Zhang X, Yuan J, Huang H, Xiang J, Li F. A novel candidate gene associated with body weight in the pacific white shrimp Litopenaeus vannamei. Front Genet. 2019, 10: 520

[46] Wang Q, Yu Y, Zhang Q, Luo Z, Zhang X, Xiang J, Li F. The polymorphism of LvMmd2 and its association with growth traits in Litopenaeus vannamei. Mar Biotechnol. 2020, 22: 564-571

[47] Warren FJ, Royall PG, Gaisford S, Butterworth PJ, Ellis PR. Binding interactions of α-amylase with starch granules: the influence of supramolecular structure and surface area. Carbohydr Polym. 2011, 86: 1038-1047

[48] Xiao H, Sun X, Lin Z, Yang Y, Zhang M, Xu Z, Liu P, Liu Z, Huang H. Gentiopicroside targets PAQR3 to activate the PI3K/AKT signaling pathway and ameliorate disordered glucose and lipid metabolism. Acta Pharm Sin B. 2022, 12: 2887-2904

[49] Xie J, Peng G, Wang M, Zhong Q, Song X, Bi J, Tang J, Feng F, Gao H, Li B. RR-1 cuticular protein TcCPR69 is required for growth and metamorphosis in Tribolium castaneum. Insect Sci. 2022, 29: 1612-1628

[50] Yamauchi T, Kamon J, Waki H, Terauchi Y, Kubota N, Hara K, Mori Y, Ide T, Murakami K, Tsuboyama-Kasaoka N, Ezaki O, Akanuma Y, Gavrilova O, Vinson C, Reitman ML, Kagechika H, Shudo K, Yoda M, Nakano Y, Tobe K, et al. . The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity. Nat Med. 2001, 7: 941-946

[51] Yamauchi T, Kamon J, Minokoshi Y, Ito Y, Waki H, Uchida S, Yamashita S, Noda M, Kita S, Ueki K, Eto K, Akanuma Y, Froguel P, Foufelle F, Ferre P, Carling D, Kimura S, Nagai R, Kahn BB, Kadowaki T. Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase. Nat Med. 2002, 8: 1288-1295

[52] Yamauchi T, Kamon J, Ito Y, Tsuchida A, Yokomizo T, Kita S, Sugiyama T, Miyagishi M, Hara K, Tsunoda M, Murakami K, Ohteki T, Uchida S, Takekawa S, Waki H, Tsuno NH, Shibata Y, Terauchi Y, Froguel P, Tobe K, et al. . Cloning of adiponectin receptors that mediate antidiabetic metabolic effects. Nature. 2003, 423: 762-769

[53] Yan Y, Lu X, Kong J, Meng X, Luan S, Dai P, Chen B, Cao B, Luo K. Molecular characterization of myostatin and its inhibitory function on myogenesis and muscle growth in Chinese shrimp, Fenneropenaeus chinensis. Gene. 2020, 758 144986

[54] Yu Y, Zhang X, Yuan J, Li F, Chen X, Zhao Y, Huang L, Zheng H, Xiang J. Genome survey and high-density genetic map construction provide genomic and genetic resources for the pacific white shrimp Litopenaeus vannamei. Sci Rep. 2015, 5: 15612

[55] Yu Y, Wang Q, Zhang Q, Luo Z, Wang Y, Zhang X, Huang H, Xiang J, Li F. Genome scan for genomic regions and genes associated with growth trait in pacific white shrimp Litopeneaus vannamei. Mar Biotechnol. 2019, 21: 374-383

[56] Yuan J, Yu Y, Zhang X, Li S, Xiang J, Li F. Recent advances in crustacean genomics and their potential application in aquaculture. Rev Aquac. 2023, 15: 1501-1521

[57] Zannini L, Delia D, Buscemi G. CHK2 kinase in the DNA damage response and beyond. J Mol Cell Biol. 2014, 6: 442-457

[58] Zhang X, Yuan J, Sun Y, Li S, Gao Y, Yu Y, Liu C, Wang Q, Lv X, Zhang X, Ma KY, Wang X, Lin W, Wang L, Zhu X, Zhang C, Zhang J, Jin S, Yu K, Kong J, et al. . Penaeid shrimp genome provides insights into benthic adaptation and frequent molting. Nat Commun. 2019, 10: 356

RIGHTS & PERMISSIONS

The Author(s)