Hostname: page-component-cd9895bd7-8ctnn Total loading time: 0 Render date: 2024-12-31T19:27:31.749Z Has data issue: false hasContentIssue false
  • English
  • Français

Intracellular Staphylococcus aureus inhibits autophagy of bovine mammary epithelial cells through activating p38α

Published online by Cambridge University Press:  24 August 2021

Run Wang ,
Wen Zhang ,
Lumei Wang ,
Na Geng ,
Xiaozhou Wang ,
Meihua Zhang ,
Jianzhu Liu ,
Yongxia Liu  and
Bo Han
Run Wang
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Wen Zhang
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Lumei Wang
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Na Geng
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Xiaozhou Wang
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Meihua Zhang
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Jianzhu Liu*
Affiliation:
Research Center for Animal Disease Control Engineering, Shandong Agricultural University, Tai'an, Shandong, China
Yongxia Liu*
Affiliation:
College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong, China
Bo Han
Affiliation:
College of Veterinary Medicine, China Agricultural University, Beijing, China
*
Author for correspondence: Yongxia Liu, Email: [email protected]; Jianzhu Liu, Email: [email protected]
Author for correspondence: Yongxia Liu, Email: [email protected]; Jianzhu Liu, Email: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Staphylococcus aureus is a common pathogen of bovine mastitis which can induce autophagy and inhibit autophagy flux, resulting in intracellular survival and persistent infection. The aim of the current study was to investigate the role of p38α in the autophagy induced by intracellular S. aureus in bovine mammary epithelial cells. An intracellular infection model of MAC-T cells was constructed, and activation of p38α was examined after S. aureus invasion. Through activating/inhibiting p38α by anisomycin/SB203580, the autophagosomes, LC3 and p62 level were analyzed by immunofluorescence and western blot. To further study the detailed mechanism of p38α, phosphorylation of ULK1ser757 was also detected. The results showed that intracellular S. aureus activated p38α, and the activation developed in a time-dependent manner. Inhibition of p38α promoted intracellular S. aureus-induced autophagy flow, up-regulated the ratio of LC3 II/I, reduced the level of p62 and inhibited the phosphorylation of ULK1ser757, whereas the above results were reversed after activation of p38α. The current study indicated that intracellular S. aureus can inhibit autophagy flow by activating p38α in bovine mammary epithelial cells.

Keywords

Autophagybovine mammary epithelial cellsmastitisp38αStaphylococcus aureus
Type
Research Article
Information
Journal of Dairy Research , Volume 88 , Issue 3 , August 2021 , pp. 293 - 301
Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press on behalf of Hannah Dairy Research Foundation

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

*

These authors contributed equally to this work.

References

Barkema, HW, Schukken, YH and Zadoks, RN (2006) Invited review: the role of cow, pathogen, and treatment regimen in the therapeutic success of bovine Staphylococcus aureus mastitis. Journal of Dairy Science 89, 1877–1895.CrossRefGoogle ScholarPubMed
Bradley, A (2002) Bovine mastitis: an evolving disease. Veterinary Journal 164, 116–128.CrossRefGoogle Scholar
Burotto, M, Chiou, VL, Lee, J-M and Kohn, EC (2014) The MAPK pathway across different malignancies: a new perspective. Cancer 120, 3446–3456.CrossRefGoogle ScholarPubMed
Chargui, A, Cesaro, A, Mimouna, S, Fareh, M, Brest, P, Naquet, P, Darfeuille-Michaud, A, Hébuterne, X, Mograbi, B and Vouret-Craviari, V (2012) Subversion of autophagy in adherent invasive Escherichia coli-infected neutrophils induces inflammation and cell death. PLoS ONE 7, e51727.CrossRefGoogle ScholarPubMed
Choi, CH, Jung, Y-K and Oh, S-H (2010) Autophagy induction by capsaicin in malignant human breast cells is modulated by p38 and extracellular signal-regulated mitogen-activated protein kinases and retards cell death by suppressing endoplasmic reticulum stress-mediated apoptosis. Molecular Pharmacology 78, 114–125.CrossRefGoogle ScholarPubMed
Contreras, GA and Rodriguez, JM (2011) Mastitis: comparative etiology and epidemiology. Journal of Mammary Gland Biology and Neoplasia 16, 339–356.CrossRefGoogle ScholarPubMed
Cuervo, AM (2004) Autophagy: in sickness and in health. Trends in Cell Biology 14, 70–77.CrossRefGoogle ScholarPubMed
Dortet, L, Mostowy, S and Cossart, P (2012) Recruitment of the major vault protein by InlK: a Listeria monocytogenes strategy to avoid autophagy. PLoS Pathogens 7, e1002168–e1002134.CrossRefGoogle Scholar
Foster, TJ (2005) Immune evasion by staphylococci. Nature Reviews Microbiology 3, 948–958.CrossRefGoogle ScholarPubMed
Fu, C, Liu, L and Li, F (2017) Acetate alters the process of lipid metabolism in rabbits. International Journal of Animal Bioscience 12, 1895–1902.CrossRefGoogle ScholarPubMed
Fugui, J, Xueyan, L, Zhengui, Y, Zhiyong, H and Yun, W (2018) Effects of forage source and particle size on feed sorting, milk production and nutrient digestibility in lactating dairy cows. Journal of Animal Pysiology and Animal Nutrition 102, 1472–1481.Google Scholar
Geng, N, Liu, K, Lu, J, Xu, Y, Wang, X, Wang, R, Liu, J, Liu, Y and Han, B (2020) Autophagy of bovine mammary epithelial cell induced by intracellular Staphylococcus aureus. Journal of Microbiology 58, 320–329.CrossRefGoogle ScholarPubMed
He, Y, She, H, Zhang, T, Xu, H, Cheng, L, Yepes, M, Zhao, Y and Mao, Z (2017) p38 MAPK inhibits autophagy and promotes microglial inflammatory responses by phosphorylating ULK1. Journal of Cell Biology 17, 315–328.Google Scholar
Hu, ZY, Yin, ZY, Lin, XY, Yan, ZG and Wang, ZH (2015) Effects of feeding fatty acid calcium and the interaction of forage quality on production performance and biochemical indexes in early lactation cow. Journal of Animal Physiology and Animal Nutrition 99, 899–904.CrossRefGoogle ScholarPubMed
Irving, AT, Mimuro, H, Kufer, TA, Lo, C, Wheeler, R, Turner, LJ, Thomas, BJ, Malosse, C, Gantier, MP, Casillas, LN, Votta, BJ, Bertin, J, Boneca, IG, Sasakawa, C, Philpott, DJ, Ferrero, RL and Kaparakis-Liaskos, M (2014) The immune receptor NOD1 and kinase RIP2 Interact with bacterial peptidoglycan on early endosomes to promote autophagy and Inflammatory signaling. Cell Host & Microbe 15, 623–635.CrossRefGoogle ScholarPubMed
Jie, Y, Zhao, X, Hu, Y, Sun, H and Kong, W (2018) Autophagy regulates the degeneration of the auditory cortex through the AMPK-mTOR-ULK1 signaling pathway. International Journal of Molecular Medicine 41, 2086–2098.Google Scholar
Kim, J and Guan, KL (2011) Regulation of the autophagy initiating kinase ULK1 by nutrients: roles of mTORC1 and AMPK. Cell Cycle 10, 1337–1338.CrossRefGoogle ScholarPubMed
Lamark, T, Svenningand, S and Johansen, T (2017) Regulation of selective autophagy: the p62/SQSTM1 paradigm. Essays in Biochemistry 61, 609–624.Google ScholarPubMed
Li, W, Zhu, J, Dou, J, She, H, Tao, K, Xu, H, Yang, Q and Mao, Z (2017) Phosphorylation of LAMP2A by p38 MAPK couples ER stress to chaperone-mediated autophagy. Nature Communications 8, 1763.CrossRefGoogle ScholarPubMed
Li, W, Li, S, Li, Y, Lin, X, Hu, Y, Meng, T, Wu, B, He, R and Feng, D (2018) Immunofluorescence staining protocols for major autophagy proteins including LC3, P62, and ULK1 in mammalian cells in response to normoxia and hypoxia. Methods in Molecular Biology 1854, 175–185.CrossRefGoogle Scholar
Liu, SQ, Wang, LY, Liu, GH, Tang, DZ, Fan, XX, Zhao, JP, Jiao, HC, Wang, XJ, Sun, SH and Lin, H (2017) Leucine alters immunoglobulin a secretion and inflammatory cytokine expression induced by lipopolysaccharide via the nuclear factor-κB pathway in intestine of chicken embryos. Animal: An International Journal of Animal Bioscience 12, 1–9.Google ScholarPubMed
Martin, F and Bhanu, S (2012) Intracellular staphylococcus aureus: live-in and let die. Frontiers in Cellular & Infection Microbiology 2, 43.Google Scholar
Mizushima, N (2010) The role of the ATG1/ULK1 complex in autophagy regulation. Current Opinion Cell Biology 22, 132–139.CrossRefGoogle ScholarPubMed
Mohamed, W, Sommer, U, Sethi, S, Domann, E and Alt, V (2014) Intracellular proliferation of S. aureus in osteoblasts and effects of rifampicin and gentamicin on S. aureus intracellular proliferation and survival. European Cell Materials 28, 258–268.CrossRefGoogle Scholar
Mostowy, S (2012) Autophagy and bacterial clearance: a not so clear picture. Cellular Microbiology 15(3), 395–402.CrossRefGoogle Scholar
Neumann, Y, Bruns, SA, Rohde, M, Prajsnar, TK and Schmitz, I (2016) Intracellular Staphylococcus aureus eludes selective autophagy by activating a host cell kinase. Autophagy 12, 2069–2084.CrossRefGoogle ScholarPubMed
Ning, C and Karantza-Wadsworth, V (2009) Role and regulation of autophagy in cancer. Biochimica et Biophysica Acta 1793, 1516–1523.Google Scholar
Niu, H, Yamaguchi, M and Rikihisa, Y (2008) Subversion of cellular autophagy by Anaplasma phagocytophilum. Cellular Microbiology 10, 593–605.CrossRefGoogle ScholarPubMed
Pankiv, S, Clausen, TH, Lamark, T, Brech, A and Johansen, T (2007) p62/SQSTM1 binds directly to ATG8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. The Journal of Biological Chemistry 282, 24131–24145.CrossRefGoogle ScholarPubMed
Radhi, OA, Davidson, S, Scott, F, Zeng, RX, Jones, DH, Tomkinson, NCO, Yu, J and Chan, EYW (2019) Inhibition of the ULK1 protein complex suppresses Staphylococcus-induced autophagy and cell death. The Journal of Biological Chemistry 294, 14289–14307.CrossRefGoogle ScholarPubMed
Ravikumar, B, Sarkar, S, Davies, JE, Futter, M, Garcia-Arencibia, M, Green-Thompson, ZW, Jimenez-Sanchez, M, Korolchuk, VI, Lichtenberg, M, Luo, S, Massey, DC, Menzies, FM, Moreau, K, Narayanan, U, Renna, M, Siddiqi, FH, Underwood, BR, Winslow, AR and Rubinsztein, DC (2010) Regulation of mammalian autophagy in physiology and pathophysiology. Physiological Reviews 90, 1383–1435.CrossRefGoogle ScholarPubMed
Schaaf, MB, Keulers, TG, Vooijs, MA and Rouschop, KM (2016) LC3/GABARAP family proteins: autophagy-(un)related functions. FASEB Journal 30, 3961–3978.CrossRefGoogle ScholarPubMed
Schnaith, A, Kashkar, H, Leggio, SA, Addicks, K, Kronke, M and Krut, O (2007) Staphylococcus aureus subvert autophagy for induction of caspase-independent host cell death. The Journal of Biological Chemistry 291, 2067–2079.Google Scholar
Schnöder, L, Hao, W, Qin, Y, Liu, S, Tomic, I, Liu, X, Fassbender, K and Liu, Y (2016) Deficiency of neuronal p38α MAPK attenuates amyloid pathology in Alzheimer disease mouse and cell models through facilitating lysosomal degradation of BACE1. The Journal of Biological Chemistry 282, 2695–2706.Google Scholar
Shahnazari, S and Brumell, JH (2011) Mechanisms and consequences of bacterial targeting by the autophagy pathway. Current Opinion in Microbiology 14, 68–75.CrossRefGoogle ScholarPubMed
Shahnazari, S, Namolovan, A, Mogridge, J, Kim, PK and Brumell, JH (2011) Bacterial toxins can inhibit host cell autophagy through cAMP generation. Autophagy 7, 957–965.10.4161/auto.7.9.16435CrossRefGoogle ScholarPubMed
Stuhr, T and Aulrich, K (2010) Intramammary infections in dairy goats: recent knowledge and indicators for detection of subclinical mastitis. Forestry 2010, 267–279.Google Scholar
Takuo, O, Jahangir, MD, Alam, N and Noda, (2018) Membrane-binding domains in autophagy. Chemistry and Physics Lipids 218, 1–9.Google Scholar
Wang, Q, Miao, Y, Xu, Y, Meng, X, Cui, W, Wang, Y, Zhu, L, Sha, Z, Wei, K and Zhu, R (2019) Taishan Pinus Massoniana pollen polysaccharide inhibits the replication of acute tumorigenic ALV-J and its associated tumor growth. Veterinary Microbiol 236, 108376.CrossRefGoogle ScholarPubMed
Webber, JL and Tooze, SA (2009) Coordinated regulation of autophagy by p38α MAPK through mATG9 and p38IP. The EMBO Journal 29, 27–40.CrossRefGoogle ScholarPubMed
Yang, Z, Wilkie-Grantham, RP, Yanagi, T, Shu, CW, Matsuzawa, S and Reed, JC (2015) ATG4B (Autophagin-1) phosphorylation modulates autophagy. The Journal of Biological Chemistry 290, 26549–26561.CrossRefGoogle ScholarPubMed
Yujuan, N, Qinqin, S, Guihua, Z, Xingpo, L, Yingli, S, Yihong, X and Sidang, L (2018) Fowl adenovirus serotype 4-induced apoptosis, autophagy, and a severe inflammatory response in liver. Veterinary Microbiology 223, 34–41.Google Scholar
Zhai, R, Dong, X, Feng, L, Li, S and Hu, Z (2019) The effect of heat stress on autophagy and apoptosis of rumen, abomasum, duodenum, liver and kidney cells in calves. Animals (Basel) 9, 854.CrossRefGoogle ScholarPubMed
Zhou, Z, Ning, C, Shuai, S, Shuhong, S and Zhizhong, C (2018) The molecular basis for host responses to Marek's disease viruses integrated with different retro-viral long terminal repeat. Poultry Science 97, 3015–3022.CrossRefGoogle ScholarPubMed
Zhu, CB, Carneiro, AM, Dostmann, WR, Hewlett, WA and Blakely, RD (2005) p38 MAPK activation elevates serotonin transport activity via a trafficking-independent, protein phosphatase 2A-dependent process. The Journal of Biological Chemistry 280, 15649–15658.CrossRefGoogle Scholar