猪ADAM10基因编码区及其剪接体克隆、序列分析与组织表达研究

doi:10.11843/j.issn.0366-6964.2017.09.003

Abstract

Abstract:

The objective of this study was to clone the coding sequence (CDS) of porcine ADAM10 gene and its splicing variant, analyze the gene structure and functions using bioinformatics methods, and investigate the mRNA levels of ADAM10 gene and its splicing variant in different tissues in porcine. According to the sequence of porcine ADAM10 gene in GenBank, the specific primers were designed for cloning the CDS of ADAM10 gene and its splicing variant using reverse transcription-PCR (RT-PCR) and T/A cloning methods. The protein structure of ADAM10 and its splicing variant were analyzed through bioinformatics methods. The mRNA expression patterns of porcine ADAM10 gene and its splicing variant in different tissues in porcine were analyzed by semi-quantitative PCR. The results showed that the complete CDS of ADAM10 gene was 2 247 bp, encoding 748 amino acids. The CDS of splicing variant, missing the sequence of exon 2-7 and partial exon 8, had a length of 1 344 bp which encoded 447 amino acids. Compared with the intact protein, the splicing variant protein missed the leading peptide sequence. However, the splicing variant protein contained signal peptide, transmembrane domain and metalloproteinase and depolymerized domains. Futhermore, the tertiary structures of the 2 proteins were identical. Sequence alignment and phylogenetic analysis showed that the intact ADAM10 protein was highly conserved among species. Semi-quantitative PCR analysis showed that the intact mRNA of ADAM10 gene was highly expressed in spleen and lowly in kidney, mammary gland, leg muscle, fallopian tube, ovary, uterus and small intestine; the mRNA of splicing variant was highly expressed in lung and lowly in spleen and stomach. Our study provides the basic materials for further studying on the structure and biological function of porcine ADAM10 gene.

CLC Number:

S828.2

CHEN Wen-da, PEI Yue, ZHOU Xiao-long, ZHAO A-yong, YANG Song-bai. Coding Sequence Cloning, Alternative Splicing, Sequence Analysis and Tissue Expression of Porcine ADAM10 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1579-1590.

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References

[1] DREYMUELLER D, PRUESSMEYER J, GROTH E, et al. The role of ADAM-mediated shedding in vascular biology[J]. Eur J Cell Biol, 2012, 91(6-7):472-485.
[2] KWON J, JEONG S M, CHOI I, et al. ADAM10 is involved in cell junction assembly in early porcine embryo development[J]. PLoS One, 2016, 11(4):e0152921.
[3] CHANTRY A, GREGSON N A, GLYNN P. A novel metalloproteinase associated with brain myelin membranes. Isolation and characterization[J]. J Biol Chem, 1989, 264(36):21603-21607.
[4] FOLKESSON M, LI C, FREBELIUS S, et al. Proteolytically active ADAM10 and ADAM17 carried on membrane microvesicles in human abdominal aortic aneurysms[J]. Thromb Haemost, 2015, 114(6):1165-1174.
[5] SEALS D F, COURTNEIDGE S A. The ADAMs family of metalloproteases:multidomain proteins with multiple functions[J]. Genes Dev, 2003, 17(1):7-30.
[6] ZHUANG J L, WEI Q L, LIN Z H, et al. Effects of ADAM10 deletion on Notch-1 signaling pathway and neuronal maintenance in adult mouse brain[J]. Gene, 2015, 555(2):150-158.
[7] FENG L Z, WANG Y J, CAI H, et al. ADAM10-Notch signaling governs the recruitment of ovarian pregranulosa cells and controls folliculogenesis in mice[J]. J Cell Sci, 2016, 129(11):2202-2212.
[8] MURAI T, MIYAUCHI T, YANAGIDA T, et al. Epidermal growth factor-regulated activation of Rac GTPase enhances CD44 cleavage by metalloproteinase disintegrin ADAM10[J]. Biochem J, 2006, 395(1):65-71.
[9] PAN Y, HAN C, WANG C L, et al. ADAM10 promotes pituitary adenoma cell migration by regulating cleavage of CD44 and L1[J]. J Mol Endocrinol, 2012, 49(1):21-33.
[10] ANDEREGG U, EICHENBERG T, PARTHAUNE T, et al. ADAM10 is the constitutive functional sheddase of CD44 in human melanoma cells[J]. J Invest Dermatol, 2009, 129(6):1471-1482.
[11] YANG K, LU L, LIU Y, et al. Increase of ADAM10 level in coronary artery in-stent restenosis segments in diabetic minipigs:high ADAM10 expression promoting growth and migration in human vascular smooth muscle cells via Notch 1 and 3[J]. PLoS One, 2013, 8(12):e83853.
[12] KANDA K, KOMEKADO H, SAWABU T, et al. Nardilysin and ADAM proteases promote gastric cancer cell growth by activating intrinsic cytokine signalling via enhanced ectodomain shedding of TNF-α[J]. EMBO Mol Med, 2012, 4(5):396-411.
[13] LOREY M B, ROSSI K, EKLUND K K, et al. Global characterization of protein secretion from human macrophages following non-canonical caspase-4/5 inflammasome activation[J]. Mol Cell Proteomics, 2017, 16(4 Suppl 1):S187-S199, doi:10.1074/mcp.M116.064840.
[14] BLACK D L. Mechanisms of alternative pre-messenger RNA splicing[J]. Annu Rev Biochem, 2003, 72:291-336.
[15] 万敬员, 张力, 叶笃筠. 选择性剪接调控机制的研究进展[J]. 国外医学分子生物学分册, 2003, 25(6):342-345.
WAN J Y, ZHANG L, YE D Y. Research advances on mechanisms of alternative splicing regulation[J]. Journal of Medical Molecular Biology, 2003, 25(6):342-345.(in Chinese)
[16] VUONG C K, BLACK D L, ZHENG S K. The neurogenetics of alternative splicing[J]. Nat Rev Neurosci, 2016, 17(5):265-281.
[17] ARDUISE C, ABACHE T, LI L, et al. Tetraspanins regulate ADAM10-mediated cleavage of TNF-α and epidermal growth factor[J]. J Immunol, 2008, 181(10):7002-7013.
[18] VINCENT B. Regulation of the α-secretase ADAM10 at transcriptional, translational and post-translational levels[J]. Brain Res Bull, 2016, 126(Pt 2):154-169.
[19] ANDERS A, GILBERT S, GARTEN W, et al. Regulation of the α-secretase ADAM10 by its prodomain and proprotein convertases[J]. FASEB J, 2001, 15(10):1837-1839.
[20] MOCHIZUKI S, OKADA Y. ADAMs in cancer cell proliferation and progression[J]. Cancer Sci, 2007, 98(5):621-628.
[21] 潘源, 丁学华. 抗癌治疗新靶标——去整合素金属蛋白酶10[J]. 国际肿瘤学杂志, 2011, 38(7):492-495.
PAN Y, DING X H. A new anti-cancer target:ADAM10[J]. Journal of International Oncology, 2011, 38(7):492-495. (in Chinese)
[22] LUNN C A, FAN X D, DALIE B, et al. Purification of ADAM 10 from bovine spleen as a TNFα convertase[J].FEBS Lett, 1997, 400(3):333-335.

Coding Sequence Cloning, Alternative Splicing, Sequence Analysis and Tissue Expression of Porcine ADAM10 Gene

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