Effects of ClpP Protease on Biofilm Formation of Enterococcus Faecalis

Overview

Journal J Appl Oral Sci

Specialty Dentistry

Date 2021 Mar 3

PMID 33656065

Citations 6

Authors

Ying Feng

Hongyuan Wang

H E Lu

Liu Yi

L I Hong

Affiliations

Soon will be listed here.

Abstract

Objectives: Enterococcus faecalis (E. faecalis), one of the main pathogens responsible for refractory periapical periodontitis and nosocomial infections, exhibits markedly higher pathogenicity in biofilms. Studies have shown that caseinolytic protease P (ClpP) is involved in biofilm formation. However, to date, few studies have investigated the role of ClpP in the survival of E. faecalis, and in enhancing biofilm formation. Therefore, we investigated the role of ClpP in the formation of E. faecalis biofilms.

Methodology: In our study, we used homologous recombination to construct clpP deleted and clpP complement strains of E. faecalis ATCC 29212. A viable colony counting method was used to analyze the growth patterns of E. faecalis. Crystal violet staining (CV) and confocal scanning laser microscopy (CLSM) were used to characterize biofilm mass formation and scanning electron microscopy (SEM) was used to observe the biofilm microstructure. Data was statistically analyzed via Student's t-test or one-way analysis of variance (ANOVA).

Results: The results exhibited altered growth patterns for the clpP deletion strains and depleted polysaccharide matrix, resulting in reduced biofilm formation capacity compared to the standard strains. Moreover, ClpP was observed to increase bioﬁlm formation in E. faecalis.

Conclusion: Our study shows that ClpP can increase bioﬁlm formation in E. faecalis and emphasizes the importance of ClpP as a potential target against E. faecalis.

Citing Articles

-mediated regulation of growth and biofilm formation in .

Zhou J, Ma Q, Liang J, Pan Y, Chen Y, Yu S Front Microbiol. 2025; 15:1507928.

PMID: 39895941 PMC: 11782273. DOI: 10.3389/fmicb.2024.1507928.

Novel function of single-target regulator NorR involved in swarming motility and biofilm formation revealed in Vibrio alginolyticus.

Chen T, Zhou X, Feng R, Shi S, Chen X, Wei B BMC Biol. 2024; 22(1):253.

PMID: 39506750 PMC: 11542441. DOI: 10.1186/s12915-024-02057-y.

Strategies and mechanisms targeting biofilms associated with endodontic infections: a comprehensive review.

Yang S, Meng X, Zhen Y, Baima Q, Wang Y, Jiang X Front Cell Infect Microbiol. 2024; 14:1433313.

PMID: 39091674 PMC: 11291369. DOI: 10.3389/fcimb.2024.1433313.

Coaggregated E. faecalis with F. nucleatum regulated environmental stress responses and inflammatory effects.

Zhou J, Yuan Z, Yang R, Liu T, Lu X, Huang W Appl Microbiol Biotechnol. 2024; 108(1):336.

PMID: 38761182 PMC: 11102388. DOI: 10.1007/s00253-024-13172-9.

Bacterial biofilm growth and perturbation by serine protease from Bacillus sp.

Yunus J, Wan Dagang W, Jamaluddin H, Jemon K, Mohamad S, Jonet M Arch Microbiol. 2024; 206(4):138.

PMID: 38436775 DOI: 10.1007/s00203-024-03857-0.

References

Zheng J, Wu Y, Lin Z, Wang G, Jiang S, Sun X . ClpP participates in stress tolerance, biofilm formation, antimicrobial tolerance, and virulence of Enterococcus faecalis. BMC Microbiol. 2020; 20(1):30. PMC: 7006429. DOI: 10.1186/s12866-020-1719-9. View

Yu M, Kim M, Rosa V, Hwang Y, Fabbro M, Sohn W . Role of extracellular DNA in Enterococcus faecalis biofilm formation and its susceptibility to sodium hypochlorite. J Appl Oral Sci. 2019; 27:e20180699. PMC: 9648955. DOI: 10.1590/1678-7757-2018-0699. View

Thomas V, Hancock L . Suicide and fratricide in bacterial biofilms. Int J Artif Organs. 2009; 32(9):537-44. DOI: 10.1177/039139880903200902. View

Hall B, Breidenstein E, de la Fuente-Nunez C, Reffuveille F, Mawla G, Hancock R . Two Isoforms of Clp Peptidase in Pseudomonas aeruginosa Control Distinct Aspects of Cellular Physiology. J Bacteriol. 2016; 199(3). PMC: 5237113. DOI: 10.1128/JB.00568-16. View

Li W, Liu H, Xu Q . Extracellular dextran and DNA affect the formation of Enterococcus faecalis biofilms and their susceptibility to 2% chlorhexidine. J Endod. 2012; 38(7):894-8. DOI: 10.1016/j.joen.2012.04.007. View

Zhang J, Hou X, Song X, Ma X, Zhao Y, Zhang S . ClpP affects biofilm formation of Streptococcus mutans differently in the presence of cariogenic carbohydrates through regulating gtfBC and ftf. Curr Microbiol. 2015; 70(5):716-23. DOI: 10.1007/s00284-015-0779-9. View

Yamaguchi M, Noiri Y, Kuboniwa M, Yamamoto R, Asahi Y, Maezono H . Porphyromonas gingivalis biofilms persist after chlorhexidine treatment. Eur J Oral Sci. 2013; 121(3 Pt 1):162-8. DOI: 10.1111/eos.12050. View

He L, Wang H, Zhang R, Li H . The regulation of Porphyromonas gingivalis biofilm formation by ClpP. Biochem Biophys Res Commun. 2018; 509(2):335-340. DOI: 10.1016/j.bbrc.2018.12.071. View

Tendolkar P, Baghdayan A, Shankar N . Putative surface proteins encoded within a novel transferable locus confer a high-biofilm phenotype to Enterococcus faecalis. J Bacteriol. 2006; 188(6):2063-72. PMC: 1428127. DOI: 10.1128/JB.188.6.2063-2072.2006. View

10.

Brotz-Oesterhelt H, Sass P . Bacterial caseinolytic proteases as novel targets for antibacterial treatment. Int J Med Microbiol. 2013; 304(1):23-30. DOI: 10.1016/j.ijmm.2013.09.001. View

11.

Michel A, Agerer F, Hauck C, Herrmann M, Ullrich J, Hacker J . Global regulatory impact of ClpP protease of Staphylococcus aureus on regulons involved in virulence, oxidative stress response, autolysis, and DNA repair. J Bacteriol. 2006; 188(16):5783-96. PMC: 1540084. DOI: 10.1128/JB.00074-06. View

12.

Foster T . Antibiotic resistance in Staphylococcus aureus. Current status and future prospects. FEMS Microbiol Rev. 2017; 41(3):430-449. DOI: 10.1093/femsre/fux007. View

13.

Zheng J, Wu Y, Lin Z, Pu Z, Yao W, Chen Z . Characteristics of and Virulence Factors Associated with Biofilm Formation in Clinical Isolates in China. Front Microbiol. 2017; 8:2338. PMC: 5705541. DOI: 10.3389/fmicb.2017.02338. View

14.

Frees D, Chastanet A, Qazi S, Sorensen K, Hill P, Msadek T . Clp ATPases are required for stress tolerance, intracellular replication and biofilm formation in Staphylococcus aureus. Mol Microbiol. 2004; 54(5):1445-62. DOI: 10.1111/j.1365-2958.2004.04368.x. View

15.

Szczepanowska K, Maiti P, Kukat A, Hofsetz E, Nolte H, Senft K . CLPP coordinates mitoribosomal assembly through the regulation of ERAL1 levels. EMBO J. 2016; 35(23):2566-2583. PMC: 5283601. DOI: 10.15252/embj.201694253. View

16.

Hill N, Zuke J, Buske P, Chien A, Levin P . A nutrient-dependent division antagonist is regulated post-translationally by the Clp proteases in Bacillus subtilis. BMC Microbiol. 2018; 18(1):29. PMC: 5889556. DOI: 10.1186/s12866-018-1155-2. View

17.

Kwon H, Kim S, Choi M, Ogunniyi A, Paton J, Park S . Effect of heat shock and mutations in ClpL and ClpP on virulence gene expression in Streptococcus pneumoniae. Infect Immun. 2003; 71(7):3757-65. PMC: 162022. DOI: 10.1128/IAI.71.7.3757-3765.2003. View

18.

Riboulet E, Verneuil N, La Carbona S, Sauvageot N, Auffray Y, Hartke A . Relationships between oxidative stress response and virulence in Enterococcus faecalis. J Mol Microbiol Biotechnol. 2007; 13(1-3):140-6. DOI: 10.1159/000103605. View

19.

Chatterjee I, Maisonneuve E, Ezraty B, Herrmann M, Dukan S . Staphylococcus aureus ClpC is involved in protection of carbon-metabolizing enzymes from carbonylation during stationary growth phase. Int J Med Microbiol. 2011; 301(4):341-6. DOI: 10.1016/j.ijmm.2010.10.002. View

20.

Liu Q, Wang X, Qin J, Cheng S, Yeo W, He L . The ATP-Dependent Protease ClpP Inhibits Biofilm Formation by Regulating Agr and Cell Wall Hydrolase Sle1 in . Front Cell Infect Microbiol. 2017; 7:181. PMC: 5430930. DOI: 10.3389/fcimb.2017.00181. View