Abstract
Hemolysin-coregulated protein (Hcp) of Salmonella enteritidis is known to be a structural and effector protein of the T6SS, but little is known about the role of Hcp in host cells. In this study, Hcp was expressed by plasmid pEGFP-N1-hcp in BHK-21 cells and the results showed that the subcellular localization of Hcp was predominantly in the cytoplasm of BHK-21 cells. When Hcp was over-expressed by transfecting plasmid pCI-neo-hcp in BHK-21 cells and mRNA sequencing was performed to analyze differentially expressed genes, the results showed a change in the expression levels of 307 mRNAs (fold change > 2, and p < 0.01). Amongst these, 125 mRNAs were up-regulated and 182 mRNAs were down-regulated. Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis showed that differentially expressed genes were enriched in tumor necrosis factor (TNF) signaling pathway, IL-17 signaling pathway, and cytokine–cytokine receptor interaction. Subsequently, we selected differentially expressed genes of TNF signaling pathway and verified the changes by real-time PCR. The results were consistent with the trend observed for the sequencing results. In conclusion, we demonstrated that Hcp of Salmonella enteritidis caused the change of mRNAs expression of TNF signaling pathway in the cytoplasm of BHK-21 cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aschtgen M-S, Bernard CS, De Bentzmann S, Lloubes R, Cascales E (2008) SciN Is an outer membrane lipoprotein required for type VI secretion in enteroaggregative Escherichia coli. J Bacteriol 190:7523–7531. https://doi.org/10.1128/jb.00945-08
Brenner FW, Villar RG, Angulo FJ, Tauxe R, Swaminathan B (2000) Salmonella nomenclature—guest commentary. J Clin Microbiol 38:2465–2467
Das S, Ray S, Ryan D, Sahu B, Suar M (2018) Identification of a novel gene in ROD9 island of salmonella enteritidis involved in the alteration of virulence-associated genes expression. Virulence 9:348–362. https://doi.org/10.1080/21505594.2017.1392428
DePaolo RW, Lathan R, Rollins BJ, Karpus WJ (2005) The chemokine CCL2 is required for control of murine gastric Salmonella enterica infection. Infection And Immunity 73:6514–6522. https://doi.org/10.1128/iai.73.10.6514-6522.2005
Echtenkamp F et al (2008) Characterization of the NleF effector protein from attaching and effacing bacterial pathogens. FEMS Microbiol Lett 281:98–107. https://doi.org/10.1111/j.1574-6968.2008.01088.x
Folkesson A, Lofdahl S, Normark S (2002) The Salmonella enterica subspecies I specific centisome 7 genomic island encodes novel protein families present in bacteria living in close contact with eukaryotic cells. Res Microbiol 153:537–545. https://doi.org/10.1016/s0923-2508(02)01348-7
Galan JE (1996) Molecular and cellular bases of Salmonella entry into host cells. Curr Top Microbiol Immunol 209:43–60
Hood RD et al (2010) A Type VI secretion system of pseudomonas aeruginosa targets, a toxin to bacteria cell. Host Microbe 7:25–37. https://doi.org/10.1016/j.chom.2009.12.007
Huang H et al (2016) Transcriptome analysis indicated that Salmonella lipopolysaccharide-induced thymocyte death and thymic atrophy were related to TLR4-FOS/JUN pathway in chicks. Bmc Genomics 17:322. https://doi.org/10.1186/s12864-016-2674-6
Jones SA, Jenkins BJ (2018) Recent insights into targeting the IL-6 cytokine family in inflammatory diseases and cancer. Nat Rev Immunol 18:773–789. https://doi.org/10.1038/s41577-018-0066-7
Kanamaru S (2009) Structural similarity of tailed phages and pathogenic bacterial secretion systems. Proc Natl Acad Sci USA 106:4067–4068. https://doi.org/10.1073/pnas.0901205106
Kapitein N, Mogk A (2013) Deadly syringes: type VI secretion system activities in pathogenicity and interbacterial competition. Curr Opin Microbiol 16:52–58. https://doi.org/10.1016/j.mib.2012.11.009
Laptev GY et al (2019) Examination of the expression of immunity genes and bacterial profiles in the caecum of growing chickens infected with salmonella enteritidis and fed a phytobiotic. Animals 9(9):615. https://doi.org/10.3390/ani9090615
Lertpiriyapong K et al (2012) Campylobacter jejuni type VI secretion system: roles in adaptation to deoxycholic acid, host cell adherence, invasion, and in vivo colonization. Plos One 7:e42842. https://doi.org/10.1371/journal.pone.0042842
Lim YT et al (2015) Extended loop region of hcp1 is critical for the assembly and function of type VI secretion system in burkholderia pseudomallei. Sci Rep 5:8235. https://doi.org/10.1038/srep08235
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Mougous JD et al (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science 312:1526–1530. https://doi.org/10.1126/science.1128393
Parras-Molto M, Campos-Laborie FJ, Garcia-Dieguez J, Rosario Rodriguez-Grinolo M, Perez-Pulido AJ (2013) Classification of protein motifs based on subcellular localization uncovers evolutionary relationships at both sequence and functional levels. BMC Bioinformatics 14:229. https://doi.org/10.1186/1471-2105-14-229
Polansky O, Seidlerova Z, Faldynova M, Sisak F, Rychlik I (2018) Protein expression in the liver and blood serum in chickens in response to Salmonella Enteritidis infection. Veterinary Immunol Immunopathol 205:10–16
Pukatzki S, Ma AT, Revel AT, Sturtevant D, Mekalanos JJ (2007) Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc Natl Acad Sci USA 104:15508–15513. https://doi.org/10.1073/pnas.0706532104
Ray A, Schwartz N, Santos MdS, Zhang J, Orth K, Salomon D (2017) Type VI secretion system MIX-effectors carry both antibacterial and anti-eukaryotic activities. EMBO Rep 18:1978–1990. https://doi.org/10.15252/embr.201744226
Rodríguez A, Rodríguez M, Córdoba JJ, Andrade MJ (2015) Design of primers and probes for quantitative real-time PCR methods. PCR Primer Design 1275:31–56
Basu C (ed) (2020) PCR primer design, 2nd edn. Humana Press, Los Angeles, pp 31–56
Schwarz S, Hood RD, Mougous JD (2010) What is type VI secretion doing in all those bugs? Trends Microbiol 18:531–537. https://doi.org/10.1016/j.tim.2010.09.001
Silva CA et al (2012) Infection of Mice by Salmonella enterica Serovar Enteritidis involves additional genes that are absent in the genome of serovar typhimurium. Infection And Immunity 80:839–849. https://doi.org/10.1128/iai.05497-11
Silverman JM, Brunet YR, Cascales E, Mougous JD (2012) Structure and regulation of the type VI secretion system. In: Gottesman S, Harwood CS, Schneewind O (eds). Annual review of microbiology doi: 10.1146/annurev-micro-121809–151619
Sun H, Kamanova J, Lara-Tejero M, Galan JE (2016) A Family of salmonella type III secretion effector proteins selectively targets the NF-kappaB signaling pathway to preserve host homeostasis. PLoS Pathog 12:e1005484. https://doi.org/10.1371/journal.ppat.1005484
Swaggerty CL, Kogut MH, Ferro PJ, Rothwell L, Pevzner IY, Kaiser P (2004) Differential cytokine mRNA expression in heterophils isolated from Salmonella-resistant and -susceptible chickens. Immunology 113:139–148. https://doi.org/10.1111/j.1365-2567.2004.01939.x
Wang S et al (2019) The ferric uptake regulator represses type VI secretion system function by binding directly to the clpV promoter in Salmonella enterica serovar Typhimurium. Infect Immun. https://doi.org/10.1128/iai.00562-19
Yim L et al (2014) Repression of flagella is a common trait in field isolates of salmonella enterica serovar dublin and is associated with invasive human infections. Infection Immunity 82:1465–1476. https://doi.org/10.1128/iai.01336-13
Acknowledgements
This research was funded by grants from Natural Science Foundation of China (Grant No. 31902225) and Anhui Province Natural Science Fund Project (Grant No. 1908085QC92).
Author information
Authors and Affiliations
Contributions
LZ and SL conceived and designed the experiments; SW and ML performed the experiments; ZL analyzed the data. LZ and SL wrote the paper. All authors critically read and contributed to the manuscript and approved the final version. All authors critically read and contributed to the manuscript and approved the final version.
Corresponding author
Ethics declarations
Conflict of interest
The authors have declared that no competing interest exists.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Zheng, L., Wang, S., Ling, M. et al. Salmonella enteritidis Hcp distribute in the cytoplasm and regulate TNF signaling pathway in BHK-21 cells. 3 Biotech 10, 301 (2020). https://doi.org/10.1007/s13205-020-02296-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-020-02296-0