Citation: | YAO Tuo, LU Jie, YE Lingtong, CHEN Huasheng, WANG Jiangyong. Molecular characterization and expression pattern analysis of a defensin (HdDef1) from small abalone (Haliotis diversicolor)[J]. South China Fisheries Science, 2019, 15(6): 1-8. DOI: 10.12131/20190045 |
Defensin is one of the most important antimicrobial peptides (AMPs) that participate in invertebrate innate immunity against invading pathogens. In the present study, a novel defensin was identified in small abalone (Haliotis diversicolor) that was denoted as HdDef1 by using RNA-seq and RACE techniques. The HdDef1 cDNA contained a 201 bp open reading frame (ORF) encoding 66 amino acids including a signal peptide of 18 amino acids and a mature peptide of 48 amino acids. The mature peptide of HdDef1 shared common features of AMPs, such as lower molecular mass, net positive charge (+1) and high hydrophobic residue ratio (45%). In addition, six cysteines in the mature peptide were arranged in the pattern of C-X16-C-X3-C-X9-C-X4-C-X1-C and stabilized the α-helix/β-sheet motif (CSαβ) with three disulfide bonds (C1-C4, C2-C5 and C3-C6) in the predicted tertiary structure. Moreover, by comparision with the similar three-dimensional structure of Anopheles gambiae defensin and phylogenetic analysis, it is suggested that HdDef1 might be a new member of the arthropod defensin family. Quantitative real-time PCR analysis reveals that HdDef1 transcripts were expressed constitutively in intestine, head, gill, hepatopancreas, mantle and foot, with the highest level in hepatopancreas. After being challenged with Vibrio harveyi, HdDef1 transcripts were induced significantly in hepatopancreas. The results indicate that HdDef1 might have an important function in host defense against invasive pathogenic bacteria, but its antimicrobial activity at protein level needs further study.
[1] |
吴宁, 陈梦玫, 王素芳. 贝类免疫机制的研究进展[J]. 药物生物技术, 2017, 24(1): 68-71.
|
[2] |
PARISI M G, VIZZINI A, TOUBIANA M, et al. Identification, cloning and environmental factors modulation of a αβ defensin from the Lessepsian invasive mussel Brachidontes pharaonis (Bivalvia: Mytilidae)[J]. Invertebr Surviv J, 2015, 12(12): 264-273.
|
[3] |
ZHANG L B, YANG D L, WANG Q, et al. A defensin from clam Venerupis philippinarum: molecular characterization, localization, antibacterial activity, and mechanism of action[J]. Dev Comp Immunol, 2015, 51(1): 29-38. doi: 10.1016/j.dci.2015.02.009
|
[4] |
孙敬敬, 刘慧慧, 周世权, 等. 一种新型贻贝抗菌肽的分离纯化及鉴定[J]. 水生生物学报, 2014, 38(3): 563-570. doi: 10.7541/2014.79
|
[5] |
GERDOL M, de MORO G, MANFRIN C, et al. Big defensins and mytimacins, new AMP families of the Mediterranean mussel Mytilus galloprovincialis[J]. Dev Comp Immunol, 2012, 36(2): 390-399. doi: 10.1016/j.dci.2011.08.003
|
[6] |
QIN C L, HUANG W, ZHOU S Q, et al. Characterization of a novel antimicrobial peptide with chiting-biding domain from Mytilus coruscus[J]. Fish Shellfish Immunol, 2014, 41(2): 362-370. doi: 10.1016/j.fsi.2014.09.019
|
[7] |
宫延斌, 秦传利, 石戈, 等. 厚壳贻贝抗菌肽mytichitin-CB的固相化学合成、复性及功能[J]. 浙江海洋大学学报(自然科学版), 2018, 37(1): 8-13.
|
[8] |
JENSSEN H, HAMILL P, HANCOCK R E. Peptide antimicrobial agents[J]. Clin Microbiol Rev, 2006, 19(3): 491-511. doi: 10.1128/CMR.00056-05
|
[9] |
燕晓翠, 杨春蕾, 姚大为, 等. 抗菌肽的国内外研究进展[J]. 天津农业科学, 2017, 23(5): 35-41. doi: 10.3969/j.issn.1006-6500.2017.05.008
|
[10] |
BACHÈRE E, ROSA R D, SCHMITT P, et al. The new insights into the oyster antimicrobial defense: cellular, molecular and genetic view[J]. Fish Shellfish Immunol, 2015, 46(1, SI): 50-64. doi: 10.1016/j.fsi.2015.02.040
|
[11] |
XU W, FAISAL M. Defensin of the zebra mussel (Dreissena polymorpha): molecular structure, in vitro expression, antimicrobial activity, and potential functions[J]. Mol Immunol, 2010, 47(11/12): 2138-2147.
|
[12] |
PENG K, WANG J H, SHENG J Q, et al. Molecular characterization and immune analysis of a defensin from freshwater pearl mussel, Hyriopsis schlegelii[J]. Aquaculture, 2012, 334: 45-50.
|
[13] |
WANG Y T, ZENG Z Y, ZHANG X H, et al. Identification and characterization of a novel defensin from Asian green mussel Perna viridis[J]. Fish Shellfish Immunol, 2018, 74: 242-249. doi: 10.1016/j.fsi.2017.12.029
|
[14] |
GONZALEZ M, GUEGUEN Y, DESSERRE G, et al. Molecular characterization of two isoforms of defensin from hemocytes of the oyster Crassostrea gigas[J]. Dev Comp Immunol, 2007, 31(4): 332-339. doi: 10.1016/j.dci.2006.07.006
|
[15] |
ZHANG Y B, CUI P F, WANG Y S, et al. Identification and bioactivity analysis of a newly identified defensin from the oyster Magallana gigas[J]. Dev Comp Immunol, 2018, 85: 177-187. doi: 10.1016/j.dci.2018.04.014
|
[16] |
王鹤, 胡宝庆, 文春根, 等. 褶纹冠蚌防御素基因特征与表达[J]. 中国水产科学, 2013, 20(6): 1188-1196.
|
[17] |
JIANG Q R, SHI L Y, KE C H, et al. Identification and characterization of Vibrio harveyi associated with diseased abalone Haliotis diversicolor[J]. Dis Aquat Organ, 2013, 103(2): 133-139. doi: 10.3354/dao02572
|
[18] |
LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method[J]. Methods, 2001, 25(4): 402-408. doi: 10.1006/meth.2001.1262
|
[19] |
张果苹, 汪桂玲, 郭诗照, 等. 背角无齿蚌抗菌肽theromacin基因cDNA全序列克隆及表达分析[J]. 水产学报, 2014, 38(5): 662-670.
|
[20] |
de ZOYSA M, NIKAPITIYA C, WHANG I I, et al. Abhisin: a potential antimicrobial peptide derived from histone H2A of disk abalone (Haliotis discus discus)[J]. Fish Shellfish Immunol, 2009, 27(5): 639-646. doi: 10.1016/j.fsi.2009.08.007
|
[21] |
ARENAS G, GUZMÁN F, CÁRDENAS C, et al. A novel antifungal peptide designed from the primary structure of a natural antimicrobial peptide purified from Argopecten purpuratus hemocytes[J]. Peptides, 2009, 30(8): 1405-1411. doi: 10.1016/j.peptides.2009.05.019
|
[22] |
TASSANAKAJON A, SOMBOONWIWAT K, AMPARYUP P. Sequence diversity and evolution of antimicrobial peptides in invertebrates[J]. Dev Comp Immunol, 2015, 48(2, SI): 324-341. doi: 10.1016/j.dci.2014.05.020
|
[23] |
BULET P, STÖCKLIN R, MENIN L. Anti-microbial peptides: from invertebrates to vertebrates[J]. Immunol Rev, 2004, 198(1): 169-184. doi: 10.1111/j.0105-2896.2004.0124.x
|
[24] |
ZHU S, GAO B, TYTGAT J. Phylogenetic distribution, functional epitopes and evolution of the CSαβ superfamily[J]. Cell Mol Life Sci, 2005, 62(19-20): 2257-2269. doi: 10.1007/s00018-005-5200-6
|
[25] |
de ZOYSA M, WHANG I, LEE Y, et al. Defensin from disk abalone Haliotis discus discus: molecular cloning, sequence characterization and immune response against bacterial infection[J]. Fish Shellfish Immunol, 2010, 28(2): 261-266. doi: 10.1016/j.fsi.2009.11.005
|
[26] |
ALLAM B, ESPINOSA E P. Bivalve immunity and response to infections: are we looking at the right place?[J]. Fish Shellfish Immunol, 2016, 53(SI): 4-12.
|
[27] |
YANG J Q, LUO J F, ZHENG H P, et al. Cloning of a big defensin gene and its response to Vibrio parahaemolyticus challenge in the noble scallop Chlamys nobilis (Bivalve: Pectinidae)[J]. Fish Shellfish Immunol, 2016, 56: 445-449. doi: 10.1016/j.fsi.2016.07.030
|
[28] |
MITTA G, VANDENBULCKE F, HUBERT F, et al. Mussel defensins are synthesised and processed in granulocytes then released into the plasma after bacterial challenge[J]. J Cell Sci, 1999, 112(23): 4233-4242.
|
[29] |
IWANAGA S, LEE B L. Recent advances in the innate immunity of invertebrate animals[J]. J Biochem Mol Biol, 2005, 38(2): 128-150.
|
[30] |
JOHANSSON M W, KEYSER P, SRITUNYALUCKSANA K, et al. Crustacean haemocytes and haematopoiesis[J]. Aquaculture, 2000, 191(1/2/3): 45-52.
|
[31] |
SEO J K, CRAWFORD J M, STONE K L, et al. Purification of a novel arthropod defensin from the American oyster, Crassostrea virginica[J]. Biochem Biophys Res Commun, 2005, 338(4): 1998-2004. doi: 10.1016/j.bbrc.2005.11.013
|
[32] |
崔旭. 太平洋牡蛎防御素在毕赤酵母中的重组表达及其抑菌活性[J]. 生物工程学报, 2019, 35(1): 91-101.
|
[33] |
GUEGUEN Y, HERPIN A, AUMELAS A, et al. Characterization of a defensin from the oyster Crassostrea gigas recombinant production, folding, solution structure, antimicrobial activities, and gene expression[J]. J Biol Chem, 2006, 281(1): 313-323. doi: 10.1074/jbc.M510850200
|
[34] |
WANG Q, ZHANG L B, YANG D L, et al. Molecular diversity and evolution of defensins in the manila clam Ruditapes philippinarum[J]. Fish Shellfish Immunol, 2015, 47(1): 302-312. doi: 10.1016/j.fsi.2015.09.008
|
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