| Citation: |
DOU Yiping, ZHAO Yan, PING Xiaokun, et al. Regulation mechanism of Mycobacterium bovis PtpA protein on NF-κB signaling pathway[J]. Journal of South China Agricultural University, 2019, 40(4): 1-7. DOI: 10.7671/j.issn.1001-411X.201809022
|
To investigate the effect of Mycobacterium bovis PtpA protein on NF-κB signaling pathway which is related to immune response, and reveal the role of PtpA protein in the body's response to immunity.
The PtpA gene eukaryotic expression vector FLAG-PtpA was transfected into HEK293T for SDS-PAGE analysis and Western blot detection. After activation of NF-κB signaling pathway, the effects of PtpA protein on NF-κB signaling pathway were investigated by dual luciferase assay and qPCR.
The eukaryotic expression vector FLAG-PtpA of PtpA gene was successfully constructed, transfected into HEK293T and analyzed by SDS-PAGE. The specific molecular band was visible with relative molecular mass of 22 000. Western blot results showed that the expression product specifically bound to the primary antibody, demonstrating that the protein was a PtpA protein. In the double luciferase assay, the ratios of the relative fluorescence intensities of firefly luciferase and Renilla luciferase in the test group and the control group were significantly different from 2 to 24 h after transfection (P<0.05). The relative fluorescence values of firefly luciferase and Renilla luciferase in the control group were 2.93 times higher than those in the experimental group 2 h after transfection, indicating that PtpA protein had a significant inhibitory effect on the early activation of NF-κB signaling pathway. qPCR results showed that the expression levels of IL-6, GM-CSF, BIRC-2 and BIRC-3 in the control group were 3.93, 3.42, 2.17 and 2.30 times respectively of those in the test group 2 h after transfection(P<0.01), and were 4.26, 3.93, 2.36 and 2.50 times respectively of those in the test group 4 h after transfection(P<0.01). These results indicated that PtpA protein had a significant inhibitory effect on NF-κB signaling pathway-associated cytokines (IL-6, GM-CSF, BIRC-2 and BIRC-3) in the early stage of immunization.
qPCR results are consistent with the results of dual luciferase assay, indicating that the effect of M. bovis PtpA on NF-κB signaling pathway mainly occurs in the early stage. This study provides a theoretical basis for the follow-up study of effective tuberculosis prevention and control drugs.
| [1] |
SMITH N H, GORDON S V, DE LA RUA-DOMENECH R, et al. Ecotypes of the Mycobacterium tuberculosis complex[J]. J Theor Biol, 2006, 239(2): 220-225. doi: 10.1016/j.jtbi.2005.08.036
|
| [2] |
DELAHAY R J, DE LEEUW A N, BARLOW A M, et al. The status of Mycobacterium bovisinfection in UK wild mammals: A review[J]. Vet J, 2002, 164(2): 90-105. doi: 10.1053/tvjl.2001.0667
|
| [3] |
MACIEL A, LOIKO M R, BUENO T S, et al. Tuberculosis in Southern Brazilian wild boars (Sus scrofa): First epidemiological findings[J]. Transbound Emerg Dis, 2018, 65(2): 518-526. doi: 10.1111/tbed.2018.65.issue-2
|
| [4] |
PHILLIPS C J C, FOSTER C R W, MORRIS P A, et al. The transmission of Mycobacterium bovis infection to cattle[J]. Res Vet Sci, 2003, 74(2): 1-15.
|
| [5] |
PUCKEN V B, KNUBBEN-SCHWEIZER G, DOPFER D, et al. Evaluating diagnostic tests for bovine tuberculosis in the southern part of Germany: A latent class analysis[J]. PLoS One, 2017, 12(6): e0179847. doi: 10.1371/journal.pone.0179847
|
| [6] |
DE KANTOR I N and RITACCO V. An update on bovine tuberculosis programmes in Latin American and Caribbean countries[J]. Vet Microbiol, 2006, 112(2/3/4): 111-118. doi: 10.1016/j.vetmic.2005.11.033
|
| [7] |
STONE M J, BROWN T J and DROBNIEWSKI F A. Human Mycobacterium bovis infections in London and Southeast England[J]. J Clin Microbiol, 2012, 50(1): 164-165. doi: 10.1128/JCM.05692-11
|
| [8] |
COWLEY S C, BABAKAIFF R and AV-GAY Y. Expression and localization of the Mycobacterium tuberculosis protein tyrosine phosphatase PtpA[J]. Res Microbiol, 2002, 153(4): 233-241. doi: 10.1016/S0923-2508(02)01309-8
|
| [9] |
CHAO Y, XING Y, CHEN Y, et al. Structure and mechanism of the phosphotyrosyl phosphatase activator[J]. Mol Cell, 2006, 23(4): 535-546. doi: 10.1016/j.molcel.2006.07.027
|
| [10] |
WANG J, LI B X, GE P P, et al. Mycobacterium tuberculosis suppresses innate immunity by coopting the host ubiquitin system[J]. Nat Immunol, 2015, 16(3): 237-245. doi: 10.1038/ni.3096
|
| [11] |
PFAFFI M W. A new mathematical model for relative quantification in real-time RT-PCR[J]. Nucleic Acids Res, 2001, 29(9): e45. doi: 10.1093/nar/29.9.e45
|
| [12] |
TONKS N K. Protein tyrosine phosphatases: From genes, to function, to disease[J]. Nat Rev Mol Cell Biol, 2006, 11(7): 833-846.
|
| [13] |
FU Y and GALAN J E. The Salmonella typhimurium tyrosine phosphatase SptP is translocated into host cells and disrupts the actin cytoskeleton[J]. Mol Microbiol, 1998, 27(2): 359-368. doi: 10.1046/j.1365-2958.1998.00684.x
|
| [14] |
GOEBEL-GOODY S M, WILSON-WALLIS E D, ROYSTON S, et al. Genetic manipulation of STEP reverses behavioral abnormalities in a fragile X syndrome mouse model[J]. Genes Brain Behav, 2012, 11(5): 586-600. doi: 10.1111/j.1601-183X.2012.00781.x
|
| [15] |
SHI L, POTTS M and KENNELLY P J. The serine, threonine, and/or tyrosine-specific protein kinases and protein phosphatases of prokaryotic organisms: A family portrait[J]. FEMS Microbiol Rev, 1998, 22(4): 229-253. doi: 10.1111/j.1574-6976.1998.tb00369.x
|
| [16] |
YONGFANG Z, KURUP P, JIAN X, et al. Genetic reduction of striatal-enriched tyrosine phosphatase (STEP) reverses cognitive and cellular deficits in an Alzheimer's disease mouse model[J]. Proc Natl Acad Sci USA, 2010, 107(44): 19014-19019. doi: 10.1073/pnas.1013543107
|
| [17] |
WONG D, BACH H, SUN J, et al. Mycobacterium tuberculosis protein tyrosine phosphatase (PtpA) excludes host vacuolar-H+-ATPase to inhibit phagosome acidification[J]. Proc Natl Acad Sci USA, 2011, 108(48): 19371-19376. doi: 10.1073/pnas.1109201108
|
| [18] |
WANG J, GE P, QIANG L, et al. The mycobacterial phosphatase PtpA regulates the expression of host genes and promotes cell proliferation[J]. Nat Commun, 2017, 8(1): 244. doi: 10.1038/s41467-017-00279-z
|
| [19] |
孟露萍, 史梦婷, 包海洋, 等. 结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响[J]. 中国畜牧兽医, 2016, 43(4): 892-898.
|
| [20] |
VOLKMAN H E, POZOS T C, ZHENG J, et al. Tuberculous granuloma induction via interaction of a bacterial secreted protein with host epithelium[J]. Science, 2010, 327(5964): 466-469. doi: 10.1126/science.1179663
|
| 1. |
邓永涛，尹苗，王聪，陈希文. 猪舍环境影响因子预测模型的研究进展. 家畜生态学报. 2025(02): 122-128 .
| |
| 2. |
刘艳昌，吴延昌，郭颖轩，李冬阳，张志霞，左现刚，李国厚. 生猪生长环境舒适度实时监测系统设计与实现. 家畜生态学报. 2024(06): 76-81+96 .
| |
| 3. |
刘丙杰，侯慕馨，冀海燕. 基于支持向量机集成的船舶舱室温湿度预测. 海军工程大学学报. 2024(03): 21-25+32 .
| |
| 4. |
朱佳明，孙彬，蒲施桦，潘学民，徐顺来，胡彬，齐仁立. 基于多环境因素分析的猪舍温湿度预测模型. 华南农业大学学报. 2024(05): 709-721 .
本站查看
| |
| 5. |
杨晓平，李雪寒，聂鹏程. 设施生猪环境调控现状研究和未来展望. 中国农业信息. 2024(02): 83-96 .
| |
| 6. |
尤德安，邬锡权，陆畅，董钊杰，曾志雄，吕恩利. 数字孪生技术在现代化猪场生产应用的研究进展. 现代农业装备. 2023(04): 2-9+70 .
| |
| 7. |
翟子鹤，陈小文，高莉平，张天柱. 基于梯度提升算法的温室黄瓜株高生长模拟. 中国农业大学学报. 2022(05): 134-145 .
| |
| 8. |
杨东轩，吴叶兰，张刚刚，刘硕. 基于边缘计算与深度学习的禽舍监测系统设计. 江苏农业科学. 2022(09): 219-225 .
| |
| 9. |
陈谦，杨涵，王宝刚，李文生，钱建平. 基于GRU神经网络模型的冷链运输温度时序预测. 农业大数据学报. 2022(01): 82-88 .
| |
| 10. |
刘炫文，刘仁鑫. 猪舍小气候环境模拟预测技术研究综述. 江西农业学报. 2022(08): 122-126 .
|
Supported by: Beijing Renhe Information Technology Co., Ltd. 
DownLoad: