Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation
Coxiella burnetii is the causative agent of the zoonotic disease Q fever. To date, the lipopolysaccharide (LPS) is the only defined and characterized virulence determinant of C. burnetii. In this study, proteome profiles of C. burnetii Nine Mile phase I (RSA 493, NMI) and its isogenic Nine Mile phas...
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Frontiers Media S.A.
2019-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.02022/full |
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| author | Jiri Dresler Jana Klimentova Petr Pajer Barbora Salovska Alena Myslivcova Fucikova Martin Chmel Gernot Schmoock Heinrich Neubauer Katja Mertens-Scholz |
| author_facet | Jiri Dresler Jana Klimentova Petr Pajer Barbora Salovska Alena Myslivcova Fucikova Martin Chmel Gernot Schmoock Heinrich Neubauer Katja Mertens-Scholz |
| author_sort | Jiri Dresler |
| collection | DOAJ |
| description | Coxiella burnetii is the causative agent of the zoonotic disease Q fever. To date, the lipopolysaccharide (LPS) is the only defined and characterized virulence determinant of C. burnetii. In this study, proteome profiles of C. burnetii Nine Mile phase I (RSA 493, NMI) and its isogenic Nine Mile phase II (RSA 439 NMII) isolate with a deep rough LPS were compared on L-929 mouse fibroblasts and in complex (ACCM-2), and defined (ACCM-D) media. Whole proteome extracts were analyzed using a label-free quantification approach. Between 659 and 1,046 C. burnetii proteins of the 2,132 annotated coding sequences (CDS) were identified in any particular experiment. Proteome profiles clustered according to the cultivation conditions used, indicating different regulation patterns. NMI proteome profiles compared to NMII in ACCM-D indicate transition from an exponential to a stationary phase. The levels of regulatory proteins such as RpoS, CsrA2, UspA1, and UspA2 were increased. Comparison of the oxidative stress response of NMI and NMII indicated that ACCM-2 represents a high oxidative stress environment. Expression of peroxidases, superoxide dismutases, as well as thioredoxins was increased for NMI. In contrast, in ACCM-D, only osmoregulation seems to be necessary. Proteome profiles of NMII do not differ and indicate that both axenic media represent similar oxidative stress environments. Deep rough LPS causes changes of the outer membrane stability and fluidity. This might be one reason for the observed differences. Proteins associated with the T4SS and Sec translocon as well as several effector proteins were detectable under all three conditions. Interestingly, none of these putatively secreted proteins are upregulated in ACCM-2 compared to ACCM-D, and L-929 mouse fibroblasts. Curiously, a higher similarity of proteomic patterns (overlapping up- and downregulated proteins) of ACCM-D and bacteria grown in cell culture was observed. Particularly, the proteins involved in a better adaptation or homeostasis in response to the harsh environment of the parasitophorous vacuole were demonstrated for NMI. This semi-quantitative proteomic analysis of C. burnetii compared axenically grown bacteria to those propagated in cell culture. |
| first_indexed | 2024-12-10T08:07:44Z |
| format | Article |
| id | doaj.art-65734de2663449e68a0f7218c1edcb53 |
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| language | English |
| last_indexed | 2024-12-10T08:07:44Z |
| publishDate | 2019-09-01 |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-65734de2663449e68a0f7218c1edcb532022-12-22T01:56:38ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-09-011010.3389/fmicb.2019.02022473742Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture CultivationJiri Dresler0Jana Klimentova1Petr Pajer2Barbora Salovska3Alena Myslivcova Fucikova4Martin Chmel5Gernot Schmoock6Heinrich Neubauer7Katja Mertens-Scholz8Military Health Institute, Prague, CzechiaFaculty of Military Health Sciences, University of Defence, Hradec Kralove, CzechiaMilitary Health Institute, Prague, CzechiaDepartment of Genome Integrity, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, CzechiaDepartment of Biology, Faculty of Science, University of Hradec Kralove, Hradec Kralove, CzechiaDepartment of Infectious Diseases, First Faculty of Medicine, Charles University and Military University Hospital Prague, Prague, CzechiaInstitute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Jena, GermanyInstitute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Jena, GermanyInstitute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Jena, GermanyCoxiella burnetii is the causative agent of the zoonotic disease Q fever. To date, the lipopolysaccharide (LPS) is the only defined and characterized virulence determinant of C. burnetii. In this study, proteome profiles of C. burnetii Nine Mile phase I (RSA 493, NMI) and its isogenic Nine Mile phase II (RSA 439 NMII) isolate with a deep rough LPS were compared on L-929 mouse fibroblasts and in complex (ACCM-2), and defined (ACCM-D) media. Whole proteome extracts were analyzed using a label-free quantification approach. Between 659 and 1,046 C. burnetii proteins of the 2,132 annotated coding sequences (CDS) were identified in any particular experiment. Proteome profiles clustered according to the cultivation conditions used, indicating different regulation patterns. NMI proteome profiles compared to NMII in ACCM-D indicate transition from an exponential to a stationary phase. The levels of regulatory proteins such as RpoS, CsrA2, UspA1, and UspA2 were increased. Comparison of the oxidative stress response of NMI and NMII indicated that ACCM-2 represents a high oxidative stress environment. Expression of peroxidases, superoxide dismutases, as well as thioredoxins was increased for NMI. In contrast, in ACCM-D, only osmoregulation seems to be necessary. Proteome profiles of NMII do not differ and indicate that both axenic media represent similar oxidative stress environments. Deep rough LPS causes changes of the outer membrane stability and fluidity. This might be one reason for the observed differences. Proteins associated with the T4SS and Sec translocon as well as several effector proteins were detectable under all three conditions. Interestingly, none of these putatively secreted proteins are upregulated in ACCM-2 compared to ACCM-D, and L-929 mouse fibroblasts. Curiously, a higher similarity of proteomic patterns (overlapping up- and downregulated proteins) of ACCM-D and bacteria grown in cell culture was observed. Particularly, the proteins involved in a better adaptation or homeostasis in response to the harsh environment of the parasitophorous vacuole were demonstrated for NMI. This semi-quantitative proteomic analysis of C. burnetii compared axenically grown bacteria to those propagated in cell culture.https://www.frontiersin.org/article/10.3389/fmicb.2019.02022/fullCoxiella burnetiiaxenic culturesemi-quantitative proteomicsoxidative stress responsetype IV B secretion systemdot/icm |
| spellingShingle | Jiri Dresler Jana Klimentova Petr Pajer Barbora Salovska Alena Myslivcova Fucikova Martin Chmel Gernot Schmoock Heinrich Neubauer Katja Mertens-Scholz Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation Frontiers in Microbiology Coxiella burnetii axenic culture semi-quantitative proteomics oxidative stress response type IV B secretion system dot/icm |
| title | Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation |
| title_full | Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation |
| title_fullStr | Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation |
| title_full_unstemmed | Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation |
| title_short | Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation |
| title_sort | quantitative proteome profiling of coxiella burnetii reveals major metabolic and stress differences under axenic and cell culture cultivation |
| topic | Coxiella burnetii axenic culture semi-quantitative proteomics oxidative stress response type IV B secretion system dot/icm |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.02022/full |
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