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Intracellular Ser/Thr/Tyr phosphoproteome of the oral commensal Streptococcus gordonii DL1
Malmö University, Faculty of Odontology (OD). Malmö University, Biofilms Research Center for Biointerfaces.ORCID iD: 0000-0001-9545-9161
Malmö University, Faculty of Odontology (OD). Malmö University, Biofilms Research Center for Biointerfaces.ORCID iD: 0000-0003-3173-7577
Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
Malmö University, Faculty of Odontology (OD). Malmö University, Biofilms Research Center for Biointerfaces.ORCID iD: 0000-0002-8183-8846
2020 (English)In: BMC Microbiology, E-ISSN 1471-2180, Vol. 20, no 1, article id 280Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: To respond and adapt to environmental challenges, prokaryotes regulate cellular processes rapidly and reversibly through protein phosphorylation and dephosphorylation. This study investigates the intracellular proteome and Ser/Thr/Tyr phosphoproteome of the oral commensal Streptococcus gordonii. Intracellular proteins from planktonic cells of S. gordonii DL1 were extracted and subjected to 2D-gel electrophoresis. Proteins in general were visualized using Coomassie Brilliant Blue and T-Rex staining. Phosphorylated proteins were visualized with Pro-Q Diamond Phosphoprotein Gel Stain. Proteins were identified by LC-MS/MS and sequence analysis.

RESULTS: In total, sixty-one intracellular proteins were identified in S. gordonii DL1, many of which occurred at multiple isoelectric points. Nineteen of these proteins were present as one or more Ser/Thr/Tyr phosphorylated form. The identified phosphoproteins turned out to be involved in a variety of cellular processes.

CONCLUSION: Nineteen phosphoproteins involved in various cellular functions were identified in S. gordonii. This is the first time the global intracellular Ser/Thr/Tyr phosphorylation profile has been analysed in an oral streptococcus. Comparison with phosphoproteomes of other species from previous studies showed many similarities. Proteins that are consistently found in a phosphorylated state across several species and growth conditions may represent a core phosphoproteome profile shared by many bacteria.

Place, publisher, year, edition, pages
BioMed Central, 2020. Vol. 20, no 1, article id 280
Keywords [en]
2DE, Oral bacteria, Phosphoproteome, Pro-Q diamond, Streptococci, Streptococcus gordonii
National Category
Dentistry
Identifiers
URN: urn:nbn:se:mau:diva-18388DOI: 10.1186/s12866-020-01944-yISI: 000573079700001PubMedID: 32928109Scopus ID: 2-s2.0-85091053882OAI: oai:DiVA.org:mau-18388DiVA, id: diva2:1470357
Available from: 2020-09-24 Created: 2020-09-24 Last updated: 2024-02-05Bibliographically approved
In thesis
1. Responses to External Cues in Oral Bacteria
Open this publication in new window or tab >>Responses to External Cues in Oral Bacteria
2023 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis investigates responses to external cues in oral bacteria on a molecular level. Paper I maps Ser/Thr/Tyr phosphorylated proteins in relation to the general proteome in an oral commensal streptococcus (Streptococcus gordonii DL1). The identified phosphoproteins were involved in various bacterial processes, several associated to dysbiosis and development of biofilm-induced disease. Comparison against phosphoproteomes of other bacteria showed many similarities. This is of interest for the identification of shared phosphorylation profiles. 

Paper II studies differences between the S. gordonii DL1 general proteomes in planktonic and biofilm growth phases, and the regulatory effects of salivary mucin MUC5B on protein expression in the biofilm cells. Regulations in protein expression between the different growth conditions provides insights in bacterial mechanisms for adaptation to the biofilm lifestyle. 

Paper III examines the regulatory roles of salivary MUC5B on biofilm attachment and metabolic output in two clinical isolates of oral commensals, S. gordonii CW and Actinomyces naeslundii CW. S. gordonii facilitated adhesion of A. naeslundii to MUC5B during early attachment. Both bacteria were also able to utilize MUC5B as a sole nutrient source during early biofilm formation, individually and synergistically in a dual species biofilm. The specific responses elicited by MUC5B in paper II-III seem to promote commensal colonization while down-regulating dysbiosis-related biofilm activities. 

Microbiological studies are often focused on dysbiosis and development of disease. However, mechanisms that promote eubiosis are equally important to understand how health can be maintained. Findings associated with responses to external cues in oral bacteria may contribute to future development of novel preventative strategies and identification of predictive biomarkers for oral health. 

Abstract [sv]

Avhandlingen undersöker orala bakteriers svar till utifrån kommande signaler på en molekylärbiologisk nivå. I delarbete I kartläggs proteiner med serin/treonin/ tyrosin fosforylering i relation till det generella proteomet hos en oral streptokock (Streptococcus gordonii DL1). De identifierade fosfoproteinerna kunde kopplas till olika bakteriella processer, varav flera av intresse ur kariessynpunkt. Jämförelse mot andra bakteriers fosfoproteom visade många likheter, vilket är av intresse för identifiering av delade fosforyleringsprofiler. 

I delarbete II undersöks skillnader i det generella proteomet hos S. gordonii DL1 mellan tillväxtfaserna planktonisk och biofilm, samt reglerande effekter av salivmucinet MUC5B på biofilmscellerna proteinuttryck. Skillnaderna i proteinuttryck mellan tillväxtfaser ger ledtrådar om bakteriernas mekanismer för anpassning till biofilmstillväxt. 

Delarbete III studeras salivmucinet MUC5Bs reglerande roll på vidhäftning och metabolism i biofilmer med kliniska isolat av S. gordonii CW samt Actinomyces naeslundii CW. S. gordonii ökade vidhäftningen av A. naeslundii till MUC5B. Båda arterna kunde också använda MUC5B som enda näringskälla under tidig biofilmsbildning, både enskilt och tillsammans. Responserna som MUC5B framkallade i biofilmerna (paper II och III) verkar främja kolonisering av kommensaler och samtidigt nedreglera kariesrelaterade aktiviteter. 

Mikrobiologiska studier fokuserar ofta på dysbiosis och sjukdomsutveckling, men mekanismer som bibehåller eubiosis är minst lika viktiga för att förstå hur oral hälsa kan främjas. Resultat kopplade till orala bakteriers svar på utifrån kommande signaler kan bidra till framtida utveckling av nya strategier för prevention och identifiering av prediktiva biomarkörer för oral hälsa. 

Place, publisher, year, edition, pages
Malmö: Malmö University Press, 2023. p. 105
Series
Malmö University Odontological Dissertations, ISSN 1650-6065
Keywords
Oral Microbiology, Oral Biofilms, Saliva, Salivary mucin MUC5B, Streptococcus, Actinomyces
National Category
Microbiology Biochemistry and Molecular Biology Cell and Molecular Biology Dentistry
Identifiers
urn:nbn:se:mau:diva-62571 (URN)10.24834/isbn.9789178774180 (DOI)978-91-7877-417-3 (ISBN)978-91-7877-418-0 (ISBN)
Public defence
2023-12-08, KL:2370, Carl Gustafs väg 34, Malmö, 09:00 (English)
Opponent
Supervisors
Projects
Foresight
Funder
Swedish Research Council, 201601994NIH (National Institutes of Health), P005403501
Available from: 2023-10-27 Created: 2023-10-26 Last updated: 2024-02-23Bibliographically approved

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Robertsson, CarolinaSvensäter, GunnelBlum, ZoltanWickström, Claes

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