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Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation

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dc.contributor.author Dresler, Jiri cze
dc.contributor.author Klimentova, Jana cze
dc.contributor.author Pajer, Petr cze
dc.contributor.author Salovska, Barbora cze
dc.contributor.author Myslivcová Fučíková, Alena cze
dc.contributor.author Chmel, Martin cze
dc.contributor.author Schmoock, Gernot cze
dc.contributor.author Neubauer, Heinrich cze
dc.contributor.author Mertens-Scholz, Katja cze
dc.date.accessioned 2019-10-17T07:28:51Z
dc.date.available 2019-10-17T07:28:51Z
dc.date.issued 2019 eng
dc.identifier.issn 1664-302X eng
dc.identifier.uri http://hdl.handle.net/20.500.12603/65
dc.description.abstract 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 NineMile phase I (RSA 493, NMI) and its isogenic NineMile 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. eng
dc.format p. 1-13 eng
dc.language.iso eng eng
dc.publisher Frontiers media eng
dc.relation.ispartof Frontiers in microbiology, volume 10, issue: SEP eng
dc.rights Pouze v rámci univerzity eng
dc.subject Coxiella burnetii eng
dc.subject axenic culture eng
dc.subject semi-quantitative proteomics eng
dc.subject oxidative stress response eng
dc.subject type IV B secretion system eng
dc.subject dot/icm eng
dc.subject Sec translocon eng
dc.subject proteome eng
dc.subject Coxiella burnetii cze
dc.subject axenická kultura cze
dc.subject semikvantitativní proteomika cze
dc.subject oxidační stresová reakce cze
dc.subject sekreční systém typ IV B cze
dc.subject dot/icm cze
dc.subject Sec translokon cze
dc.subject proteom cze
dc.title Quantitative Proteome Profiling of Coxiella burnetii Reveals Major Metabolic and Stress Differences Under Axenic and Cell Culture Cultivation eng
dc.title.alternative Kvantitativní analýza proteomu Coxiella burnetii odhaluje velké rozdíly v metabolismu a stresové odpovědi při porovnání axenické kultivace a kultivace v a buněčných kulturách cze
dc.type article eng
dc.identifier.obd 43875421 eng
dc.identifier.doi 10.3389/fmicb.2019.02022 eng
dc.description.abstract-translated 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 NineMile phase I (RSA 493, NMI) and its isogenic NineMile 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. cze
dc.publicationstatus postprint eng
dc.peerreviewed yes eng


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