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The Pseudogene BMEA_B0173 Deficiency in Brucella melitensis Contributes to M-epitope Formation and Potentiates Virulence in a Mice Infection Model

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Abstract

It is widely acknowledged that pseudogenes play important roles in bacterial diversification and evolution and participate in gene regulation and RNA interference (RNAi). However, the function of most pseudogenes in Brucella spp remains poorly understood, warranting further studies.To comprehensively analyze the function of the pseudogenes BMEA_B0173 in Brucella melitensis strain 63/9, a BMEA_B0173 in-frame deleted mutant strain was constructed. Then, the phenotypes of the mutant strain, such as growth characteristics and bacterial virulence, were assessed in mice infection models. Finally, iTRAQ analysis was performed to investigate the gene expression profile affected by the pseudogenes BMEA_B0173. In this study, we found that BMEA_B0173 deletion exhibited increased agglutination with M monospecific sera. In a mouse model of chronic infection, the BMEA_B0173 deletion strain displayed increased colonization in the spleen compared to the wild-type pathogen. The iTRAQ assay revealed that 252 proteins were differentially expressed between the BMEA_B0173 deletion and the wild-type strains. In addition, deletion of BMEA_B0173 significantly increased the expression of proteins involved in the denitrification pathway, iron metabolism, and several transcriptional regulators, which might cause increased virulence of the mutant strain. In conclusion, this study preliminary uncovered the function of the pseudogene BMEA_B0173 in Brucella melitensis 63/9 and provided novel insights for studying the pathogenesis of Brucella strains.

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Data Availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the iProX partner repository with the dataset identifier PXD036210.

References

  1. Ahmed W, Zheng K, Liu Z (2016) Establishment of chronic infection: Brucella’s stealth strategy. Front Cell Infect Mi 6:30. https://doi.org/10.3389/fcimb.2016.00030

    Article  CAS  Google Scholar 

  2. Corbel M (1997) Brucellosis: an overview. Emerg Infect Dis 3(2):213–221. https://doi.org/10.3201/eid0302.970219

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Suárez-Esquivel M, Chaves-Olarte E, Moreno E, Guzmán-Verri C (2020) Brucella genomics: macro and micro evolution. Int J Mol Sci 21(20):7749. https://doi.org/10.3390/ijms21207749

    Article  CAS  PubMed Central  Google Scholar 

  4. Wattam AR, Williams KP, Snyder EE, Almeida NF, Shukla M, Dickerman AW, Crasta OR, Kenyon R, Lu J, Shallom JM (2009) Analysis of ten brucella genomes reveals evidence for horizontal gene transfer despite a preferred intracellular lifestyle. J Bacteriol 191:3569

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Suárez-Esquivel M, Baker KS, Ruiz-Villalobos N, Hernández-Mora G, Barquero-Calvo E, González-Barrientos R, Castillo-Zeledón A, Jiménez-Rojas C, Chacón-Díaz C, Cloeckaert A, Chaves-Olarte E, Thomson NR, Moreno E, Guzmán-Verri C (2017) Brucella genetic variability in wildlife marine mammals populations relates to host preference and ocean distribution. Genome Biol Evol 9(7):1901–1912. https://doi.org/10.1093/gbe/evx137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Dagan T, Blekhman R, Graur AD (2006) The “domino theory” of gene death: Gradual and mass gene extinction events in three lineages of obligate symbiotic bacterial pathogens. Mol Biol Evol 23(2):310–316

    Article  CAS  PubMed  Google Scholar 

  7. Goodhead I, Darby AC (2015) Taking the pseudo out of pseudogenes. Curr Opin Microbiol 23:102–109. https://doi.org/10.1016/j.mib.2014.11.012

    Article  CAS  PubMed  Google Scholar 

  8. Sasidharan R, Gerstein M (2008) Protein fossils live on as RNA. Nature 453(7196):729–731. https://doi.org/10.1038/453729a

    Article  CAS  PubMed  Google Scholar 

  9. Balakirev ES, Ayala FJ (2003) Pseudogenes: are they “Junk” or functional DNA? Annu Rev Genet 37(1):123–151

    Article  CAS  PubMed  Google Scholar 

  10. Liu Y, Dong H, Peng X, Gao Q, Jiang H, Xu G, Qin Y, Niu J, Sun S, Li P, Ding J, Chen R (2019) RNA-seq reveals the critical role of Lon protease in stress response and Brucella virulence. Microb Pathogenesis 130:112–119. https://doi.org/10.1016/j.micpath.2019.01.010

    Article  CAS  Google Scholar 

  11. Stranahan LW, Khalaf OH, Garcia-Gonzalez DG, Arenas-Gamboa AM (2019) Characterization of Brucella canis infection in mice. PLoS ONE 14(6):e218809. https://doi.org/10.1371/journal.pone.0218809

    Article  CAS  Google Scholar 

  12. Varesio LM, Willett JW, Fiebig A, Crosson S (2019) A carbonic anhydrase pseudogene sensitizes select brucella lineages to low CO2 tension. J Bacteriol 201(22):e509–e519

    Article  Google Scholar 

  13. Bialer MG, Ferrero MC, Delpino MV, Ruiz-Ranwez V, Zorreguieta A (2021) Adhesive functions or pseudogenization of type va autotransporters in brucella species. Front Cell Infect Mi 11:607610

    Article  CAS  Google Scholar 

  14. Zygmunt MS, Blasco JM, Letesson J, Cloeckaert A, Moriyon I (2009) DNA polymorphism analysis of Brucella lipopolysaccharide genes reveals marked differences in O-polysaccharide biosynthetic genes between smooth and rough Brucella species and novel species-specific markers. Bmc Microbiol 9(1):92. https://doi.org/10.1186/1471-2180-9-92

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Vemulapalli R, He Y, Buccolo LS, Boyle SM, Sriranganathan N, Schurig GG (2000) Complementation of Brucella abortus RB51 with a functional wboA gene results in O-Antigen synthesis and enhanced vaccine efficacy but no change in rough phenotype and attenuation. Infect Immun 68(7):3927–3932. https://doi.org/10.1128/IAI.68.7.3927-3932.2000

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wang X, Wang Y, Ma L, Zhang R, De Y, Yang X, Wang C, Wu Q (2015) Development of an improved competitive ELISA based on a monoclonal antibody against lipopolysaccharide for the detection of bovine brucellosis. Bmc Vet Res 11(1):118. https://doi.org/10.1186/s12917-015-0436-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Stranahan LW, Arenas-Gamboa AM (2021) When the going gets rough: the significance of brucella lipopolysaccharide phenotype in host–pathogen interactions. Front Microbiol 12:713157. https://doi.org/10.3389/fmicb.2021.713157

    Article  PubMed  PubMed Central  Google Scholar 

  18. Lapaque N, Moriyon I, Moreno E, Gorvel J (2005) Brucella lipopolysaccharide acts as a virulence factor. Curr Opin Microbiol 8(1):60–66. https://doi.org/10.1016/j.mib.2004.12.003

    Article  CAS  PubMed  Google Scholar 

  19. Fernandez-Prada CM, Zelazowska EB, Nikolich M, Hadfield TL, Roop RM, Robertson GL, Hoover DL (2003) Interactions between Brucella melitensis and human phagocytes: bacterial surface O-Polysaccharide inhibits phagocytosis, bacterial killing, and subsequent host cell apoptosis. Infect Immun 71(4):2110–2119. https://doi.org/10.1128/IAI.71.4.2110-2119.2003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Gross A, Spiesser S, Terraza A, Rouot B, Caron E, Dornand J (1998) Expression and bactericidal activity of nitric oxide synthase in Brucella suis -infected murine macrophages. Infect Immun 66(4):1309–1316. https://doi.org/10.1128/IAI.66.4.1309-1316.1998

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Loisel-Meyer S, Jimenez D, Basseres E, Dornand J, Kohler S, Liautard JP, Jubier-Maurin V (2006) Requirement of norD for brucella suis virulence in a murine model of in vitro and in vivo infection. Infect Immun 74(3):1973–1976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Roop RM (2012) Metal acquisition and virulence in Brucella. Anim Health Res Rev 13(1):10–20. https://doi.org/10.1017/S1466252312000047

    Article  PubMed  PubMed Central  Google Scholar 

  23. Paulley JT, Anderson ES, Roop RM (2007) Brucella abortus requires the heme transporter BhuA for maintenance of chronic infection in BALB/c mice. Infect Immun 75(11):5248–5254. https://doi.org/10.1128/IAI.00460-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Bellaire BH, Elzer PH, Hagius S, Walker J, Baldwin CL, Roop RM (2003) Genetic organization and Iron-responsive regulation of the Brucella abortus 2,3-dihydroxybenzoic acid biosynthesis operon, a cluster of genes required for wild-type virulence in pregnant cattle. Infect Immun 71(4):1794–1803. https://doi.org/10.1128/IAI.71.4.1794-1803.2003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Elhassanny AEM, Anderson ES, Menscher EA, Roop RM (2013) The ferrous iron transporter FtrABCD is required for the virulence of Brucella abortus 2308 in mice. Mol Microbiol 88(6):1070–1082. https://doi.org/10.1111/mmi.12242

    Article  CAS  PubMed  Google Scholar 

  26. Almirón MA, Ugalde RA (2010) Iron homeostasis in Brucella abortus: the role of bacterioferritin. J Microbiol 48(5):668–673. https://doi.org/10.1007/s12275-010-0145-3

    Article  CAS  PubMed  Google Scholar 

  27. Xiong X, Li B, Zhou Z, Gu G, Li M, Liu J, Jiao H (2021) The VirB system plays a crucial role in brucella intracellular infection. Int J Mol Sci 22(24):13637. https://doi.org/10.3390/ijms222413637

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Sun YH, Rolan HG, Hartigh AD, Sondervan D, Tsolis RM (2005) Brucella abortus VirB12 is expressed during infection but is not an essential component of the type IV secretion system. Infect Immun 73(9):6048–6054

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Shanghai Personal Biotechnology Cp. Ltd. for their support for bioinformatics analysis with their Analysis Platform.

Funding

This work was supported by the National Key Research and Development Program of China (NO.2016YFD0500902) and Natural Science Foundation of China(NO.31902310).

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Author notes

  1. Ge Zhang and Hao Dong have contributed equally to this work.

Authors and Affiliations

  1. College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, 212018, China

    Ge Zhang & Xingjia Shen

  2. Insititute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Ge Zhang, Hui Jiang & Jiabo Ding

  3. Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control, Beijing, China

    Hao Dong

  4. National Reference Laboratory for Animal Brucellosis, China Institute of Veterinary Drug Control, Beijing, China

    Yu Feng, Jiali Sun, Xin Wang, Minghe Liu, Xiaowei Peng, Yinghui Zhang, Xiaoqian Zhang & Liangquan Zhu

  5. College of Animal Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao, China

    Tonglei Wu

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Contributions

JD and XS contributed to the study conception and design. Material preparation, data collection, and analyses were performed by GZ, YF, HJ, TW, XW, XP, YZ, XZ, and LZ. The first draft of the manuscript was written by GZ, JS, ML, and HD, and all the authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Jiabo Ding or Xingjia Shen.

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Conflict of interest

The authors declare no conflict of interest, financial or otherwise.

Ethical Approval

This study was approved by the Animal Ethics Committee of the China Institute of Veterinary Drug Control.

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Not applicable.

Research Involving in Animal Participants

Mice experiments were strictly performed in accordance with the Experimental Animal Regulation Ordinances set by the China National Science and Technology Commission.

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Supplementary Information

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Supplementary file1 (TIF 605 kb)—Figure S1. Pathway enrichment analysis with ClueGO for the ten hub genes

284_2022_3078_MOESM2_ESM.tif

Supplementary file2 (TIF 3534 kb)—Figure S2. The enriched pathways of 10 genes (different colors represented different pathways)

Supplementary file3 (DOCX 13 kb)

Supplementary file4 (DOCX 14 kb)

Supplementary file5 (DOCX 27 kb)

Supplementary file6 (DOCX 2632 kb)

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Zhang, G., Dong, H., Feng, Y. et al. The Pseudogene BMEA_B0173 Deficiency in Brucella melitensis Contributes to M-epitope Formation and Potentiates Virulence in a Mice Infection Model. Curr Microbiol 79, 378 (2022). https://doi.org/10.1007/s00284-022-03078-y

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