- Email: [email protected]
Lonely planet
Accepted Manuscript Lonely Planet Multicellular organisms Sophia Häfner, PhD, Postdoctoral fellow PII:
S1286-4579(16)00005-8
DOI:
10.1016/j.micinf.2015.12.007
Reference:
MICINF 4362
To appear in:
Microbes and Infection
Received Date: 23 December 2015
Please cite this article as: S. Häfner, Lonely Planet Multicellular organisms, Microbes and Infection (2016), doi: 10.1016/j.micinf.2015.12.007. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Lonely Planet Multicellular organisms *
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Exploring foreign countries demands a certain amount of preparation and a wellchosen arsenal of items to bring along, ranging from sun cream to earmuffs. William O. Douglas, Associate Justice of the Supreme Court of the United States from 1939 to 1975, decided at the end of the 50’s to have a road-trip through Pakistan, Afghanistan, Iran and Iraq to Istanbul. When asked how he would deal with eventual breakdowns and repair need of his vehicle, he confidently replied that he was also taking his wife along for this purpose. And indeed, the annual compilation of National Geographic form 1958 shows several pictures of Mercedes Douglas happily tinkering around with the car [2]. Entering the host organism is also quite an adventure for any bacterium, which will suddenly be confronted with exotic meals, elevated temperatures, fighting for the deck chairs in form of various epithelial cells and eventually being bothered by the border control alias the immune system. While the German tourist expresses sandals and white socks at the beach, the provident pathogen in turn will express a collection of proteins allowing it to deal with different challenges. However, while we know much about the clichés concerning fellow tourists, very little is now about what bacteria look like once they have made themselves at home in the host organism [3,4], despite the common agreement that ex vivo studies are unable to sufficiently mimic both the complexity of the host cell populations [5,6] and the coexistence of different, non-clonal bacterial populations at various stages of infection [7].
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“Looking like” is to be taken literally, in the sense of the proteome, which makes rather sense. Although proteins received some serious competition from non-coding RNAs over the last 15 years in terms of attention, they remain the final, most concrete output of the genetic machinery that the host has to handle [4]. The in vivo proteome will tell which enzymes are on duty and thus possible targets for inhibitors and which peptides are displayed on the bacterial surface and thus of use for vaccine development [4,5,7,8]. It is also one of the most complicated –omics to get one’s hands on. The main problem lies in the paucity of pathogen material against the overwhelming amount of host proteins [3,4,6]. Unlike desoxyribonucleic colleagues, there is no PCR to amplify sparse peptide material. For now, a minimum amount of 106 bacteria is required for satisfactory results, and usually bacteria have first to be sorted from host cells by centrifugation, immunomagnetic separation (IMS) or fluorescence-activated cells sorting (FACS) [3,4]. This introduces a certain preparation bias and prevents extensive prefractionation which potentially leads to a lack of detection of less abundant proteins, and to the loss of secreted or loosely membrane-bound material. Subsequently, the proteins are identified either by gelbased approaches or mass spectroscopy (MS) shot gun techniques, none of which is either easy to perform or to analye [4,7].
* Article highlight based on “Dynamic niche-specific adaptations in Neisseria meningitides during infection” by Yan Liu et al. [1].
ACCEPTED MANUSCRIPT
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So why not simply stick to transcriptomics? High-throughput techniques such as RNAseq and the drop in their cost have rendered whole-sample transcriptome analyses close to trivial. Nonetheless, first, mRNA and protein aren’t necessarily at a one to one ratio [7], and second, neither genes nor RNA always allow a correct prediction of the final subcellular location of a peptide and finally, peptides can acquire post-transcriptional modifications (PTM) which may play major roles namely on the cell surface [4,8].
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EP
TE D
M AN U
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Yan Liu and colleagues as well as Neisseria meningitidis [1] hereby join the yet rather small community of groups who attempted to establish the portrait of various pathogens in different in vivo models over the last six years. The pioneer studies took place in 2009 and 2010. Piper et al. looked at Shigella dysenteriae in gnotobiotic piglets [5]. while Kruh et al. disassembled Mycobacterium tuberculosis in a guinea pig model [7]. All together, some common features start to emerge from the different studies. While every bacterial species and even strain expresses indeed an in vivo-specific collection of proteins, they can nevertheless be assigned into a handful of functional classes. Stress resistance is one of them, unsurprisingly. Molecular chaperones and global stress proteins such as GroEL are always part of the team [3,5,6], other factors are highly indicative of the specific type of challenge the pathogen is facing – nitric stress caused by macrophages [7], oxidative and acid stress in the intestinal lumen [5] and osmotic stress in the lymph nodes [3]. Some unstudied proteins, like GadB in S. dysenteriae could in this way be related to the stress response pathways [5]. The in vivo proteome reveals also very clearly the precise metabolic adaption of the pathogen [6,7]. Shigella for examples switches to an anaerobic energy metabolism inside the host and activates numerous glycolytic enzymes [5]. Similarly, the transport of various ions - iron, phosphate, copper - seems to be a major concern to the invading bacteria, proving that the adjustment of cation levels is critical for their survival. Metal-cation transporting ATPases, efflux pumps and other transporters are a common finding of several studies and M. tuberculosis’ involvement with copper might serve as a future drug target [7]. Adhesion to host cells and fellow bacteria represents another central occupation of the exploring pathogen. In N. meningitidis, different functions, such as adhesion to epithelial cells, auto-aggregation, reshaping of the host cell membrane or DNA exchange, require different, precise number of pili per bacterium. Modest decreases in pili-related proteins and thus in the “piliation index” elicit strong phenotypes, proving that determining exact protein levels can sometimes be an important clue to function [9]. Finally, an increase of ribosomal proteins and components of the translation machinery probably hint replication stages [5,6]. Interestingly, in N. meningitidis, whose main evolutionary driving force is the incorporation of exogenous DNA and homologous recombination, genes linked to the previously described functional classes, lile pili formation and adhesion, represent recombination hotspots [8]. Apart from these predictable categories, the in vivo studies unearthed some surprises, starting with a considerable number of proteins with undefined functions, which, considering the strong selection pressure exerted on pathogens, are likely to be in charge of important tasks and thus worth a closer look [4,6]. On the contrary, Shiga toxin SD1, thought to be a crucial virulence factor, turned out not to be expressed in the host [5].
ACCEPTED MANUSCRIPT
RI PT
Another important finding is that place and time matter. Liu et al. demonstrate that different locations in the same host organism induce different protein profiles [1], Kruh and colleagues have analyzed M. tuberculosis at 30 and 90 days of infection in the lungs of guinea pigs and found little overlap between the two time points. This allows many interpretations, from the emergence of drug-tolerant steady bacterial population to the drying up of the bacteria’s favorite carbon nutrients [7]. Needless to say that this adds a layer of complexity to the in vivo studies. Bacterial populations at different stages of infection probably coexist in one host organism and adapt independently to different organs and locations. The interpretation of obtained data thus request a careful spatiotemporal dissection [3,5,6].
TE D
M AN U
SC
Nevertheless, a recent study has decided to take the pathogen in vivo proteome a daring step further by looking exclusively at the surface-exposed bacterial and adherent host proteins. Rees et al. applied a technique termed “membrane shaving”, consisting in the enzymatic cleavage and subsequent identification of membraneanchored peptides, to Corynebacterium pseudotuberculosis from sheep lymph nodes [3]. The scarcity and hydrophobicity of numerous membrane proteins has hampered their detection and quantification by gel-based and MS techniques, when they are not already lost during purification, despite the fact that they are in pole position to identify unknown players of host-pathogen interactions and to provide protective antigens [4]. The study revealed about 250 bacterial and 50 host proteins. To the list of surprises adds the statement that a substantial amount of identified peptides lacks a predicted surface feature, similar to observations of bacterial exosomes, also missing known secretion signals, proving once more that the subcellular location can’t always been inferred from the genomic sequence [3,10].
AC C
EP
All in all, cell culture and in vitro studies, attempting to mimic various stress conditions, provided us with quite interesting insights into various facets of hostpathogen interactions and stress responses by the bacterium, but the big picture was still missing [6,7]. Also, due to redundancies and the complex interplay of thousands of factors, the shift to investigate a system, such as the proteome, as a whole, has greatly improved our understanding of the many faces of a bacterium [4]. In vivo analyses are the ultimate clue to the elementary physiological state of the pathogen, a glimpse at what the enemy truly looks like [6,7]. Surely, major improvements are still required. In the close future, efforts should focus on enhancing the coverage of some highly interesting subfractions of the bacterial proteome, including the “secretome”, membrane bound peptides and glycan modifications of the latter as well as pathogencontaining host cell compartments such as the phagolysosome [4,7,8]. There is a minor drawback though. As the discussed in vivo studies usually involve euthanatizing the test subject and grinding their organs, they tend to be rather animal models than humans, thus not a hundred percent representative to what will ultimately go on in the human host. Lonely planet – H. sapiens sapiens will have to wait for a while.
ACCEPTED MANUSCRIPT 1. Background
• •
Neisseria meningitidis is a commensal meningococcus colonizing the nasopharynx of healthy humans Under unclear circumstances, it can become invasive and cause septicemia or meningitis The incidence is highest in sub-Saharan Africa (1000-100 000 persons/year), affected by periodic epidemics during the dry season
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2. In a nutshell
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Neisseria meningitidis displays distinct protein expression profiles at different infection sites in the mouse Protein abundance is high in the cerebrospinal fluid compared to blood and nasal mucosa Glutamate metabolism-associated protein expression increases in all isolates, underscoring the importance of this pathway in pathogenicity In blood, the expression of protein biogenesis components and pili proteins is enhanced PilQ favors bacterial adhesion and impedes uptake by macrophages, while GroEL is important for bacterial survival in high concentrations of human serum Deletion of NMC0101, GroEL and GuaB renders the bacterium more resistant to phagocytosis and deletion of PilQand GuaB improves survival in human serum
References
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Insert Cartoon nr. 26
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[1] Liu Y, Zhang D, Engström Å, Merényi G, Hagner M, Yanh H et al. Dynamic niche-sepcific adaptations in Neisseria meningitdis during infection. Microbes Infect 2015:http://dx.doi.org/10.1016/j.micinf.2015.09.025. [2] Winnemuth M. Das große Los: Wie ich bei Günther Jauch eine halbe Million gewann und einfach losfuhr (2012). Knaus Verlag. ISBN: 978-3-8135-0504-7. [3] Rees MA, Kleifeld O, Crellin PK, Ho B, Stinear TP, Smith AI et al. Proteomic characterization of a natural host-pathogen interaction: repertoire of in vivo expressed bacterial and host surface-associated proteins. J Proteome Res 2015;14(1):120-132. [4] Schmidt F, Völker U. Proteome analysis of host-pathogen interactions: investigation of pathogen responses to the host cell environment. Proteomics 2011;11815):3203-3211.
ACCEPTED MANUSCRIPT [5] Pieper R, Zhang Q, Parmar PP, Huang ST, Clark DJ, Alami H. The Shigella dysenteriae serotype 1 proteome, profiled in the host intestinal environment, reveals major metabolic modifications and increased expression of invasive proteins. Proteomics 2009;9(22):5029-5045.
RI PT
[6] Páková, Brychta M, Strašková A, Schmidt M, Macela A, Stulík J. Comparative proteome profiling of host-pathogen interactions: insights into the adaption mechanisms of Francisella tularensis in the host cell environment. Appl Microbiol Biotechnol 2013;97(23):10103-10115. [7] Kruh NA, Troudt J, Izzo A, Prenni J, Dobos KM. Portrait of a pathogen: the Mycobacterium tuberculosis proteome in vivo. PLoS One 2010;5(11):e13938.
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[8] Lamelas A, Harris SR, Röltgen K, Dangy JP, Hauser J, Kigsley RA et al. Emergence of a new epidemic Neisseria meningitidis serogroup A clone in the African meningitis belt: high-resolution picture of genomic changes that mediate immune evasion. MBio 2014;5(5):e01974-019714.
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[9] Imhaus AF, Duménil G. The number of Neisseria meningitidis type IV pili determines host cell interaction. EMBO J 2014;33(16):1767-1783.
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[10] Cabrita P, Trigo MJ, Ferreira RB, Brito L. Is the exoproteome important for bacterial pathogenesis? Lessons learned from interstrain exoprotein diversity in Llisteria monocytogenes grown at different temperatures. OMICS 2014;18(9):553569.
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Sophia Häfner, PhD: [email protected] Postdoctoral fellow, University of Copenhagen, BRIC Biotech Research & Innovation Centre, Lund Group, 2200 Copenhagen, Denmark
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ACCEPTED MANUSCRIPT
S1286-4579(16)00005-8
DOI:
10.1016/j.micinf.2015.12.007
Reference:
MICINF 4362
To appear in:
Microbes and Infection
Received Date: 23 December 2015
Please cite this article as: S. Häfner, Lonely Planet Multicellular organisms, Microbes and Infection (2016), doi: 10.1016/j.micinf.2015.12.007. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Lonely Planet Multicellular organisms *
M AN U
SC
RI PT
Exploring foreign countries demands a certain amount of preparation and a wellchosen arsenal of items to bring along, ranging from sun cream to earmuffs. William O. Douglas, Associate Justice of the Supreme Court of the United States from 1939 to 1975, decided at the end of the 50’s to have a road-trip through Pakistan, Afghanistan, Iran and Iraq to Istanbul. When asked how he would deal with eventual breakdowns and repair need of his vehicle, he confidently replied that he was also taking his wife along for this purpose. And indeed, the annual compilation of National Geographic form 1958 shows several pictures of Mercedes Douglas happily tinkering around with the car [2]. Entering the host organism is also quite an adventure for any bacterium, which will suddenly be confronted with exotic meals, elevated temperatures, fighting for the deck chairs in form of various epithelial cells and eventually being bothered by the border control alias the immune system. While the German tourist expresses sandals and white socks at the beach, the provident pathogen in turn will express a collection of proteins allowing it to deal with different challenges. However, while we know much about the clichés concerning fellow tourists, very little is now about what bacteria look like once they have made themselves at home in the host organism [3,4], despite the common agreement that ex vivo studies are unable to sufficiently mimic both the complexity of the host cell populations [5,6] and the coexistence of different, non-clonal bacterial populations at various stages of infection [7].
AC C
EP
TE D
“Looking like” is to be taken literally, in the sense of the proteome, which makes rather sense. Although proteins received some serious competition from non-coding RNAs over the last 15 years in terms of attention, they remain the final, most concrete output of the genetic machinery that the host has to handle [4]. The in vivo proteome will tell which enzymes are on duty and thus possible targets for inhibitors and which peptides are displayed on the bacterial surface and thus of use for vaccine development [4,5,7,8]. It is also one of the most complicated –omics to get one’s hands on. The main problem lies in the paucity of pathogen material against the overwhelming amount of host proteins [3,4,6]. Unlike desoxyribonucleic colleagues, there is no PCR to amplify sparse peptide material. For now, a minimum amount of 106 bacteria is required for satisfactory results, and usually bacteria have first to be sorted from host cells by centrifugation, immunomagnetic separation (IMS) or fluorescence-activated cells sorting (FACS) [3,4]. This introduces a certain preparation bias and prevents extensive prefractionation which potentially leads to a lack of detection of less abundant proteins, and to the loss of secreted or loosely membrane-bound material. Subsequently, the proteins are identified either by gelbased approaches or mass spectroscopy (MS) shot gun techniques, none of which is either easy to perform or to analye [4,7].
* Article highlight based on “Dynamic niche-specific adaptations in Neisseria meningitides during infection” by Yan Liu et al. [1].
ACCEPTED MANUSCRIPT
RI PT
So why not simply stick to transcriptomics? High-throughput techniques such as RNAseq and the drop in their cost have rendered whole-sample transcriptome analyses close to trivial. Nonetheless, first, mRNA and protein aren’t necessarily at a one to one ratio [7], and second, neither genes nor RNA always allow a correct prediction of the final subcellular location of a peptide and finally, peptides can acquire post-transcriptional modifications (PTM) which may play major roles namely on the cell surface [4,8].
AC C
EP
TE D
M AN U
SC
Yan Liu and colleagues as well as Neisseria meningitidis [1] hereby join the yet rather small community of groups who attempted to establish the portrait of various pathogens in different in vivo models over the last six years. The pioneer studies took place in 2009 and 2010. Piper et al. looked at Shigella dysenteriae in gnotobiotic piglets [5]. while Kruh et al. disassembled Mycobacterium tuberculosis in a guinea pig model [7]. All together, some common features start to emerge from the different studies. While every bacterial species and even strain expresses indeed an in vivo-specific collection of proteins, they can nevertheless be assigned into a handful of functional classes. Stress resistance is one of them, unsurprisingly. Molecular chaperones and global stress proteins such as GroEL are always part of the team [3,5,6], other factors are highly indicative of the specific type of challenge the pathogen is facing – nitric stress caused by macrophages [7], oxidative and acid stress in the intestinal lumen [5] and osmotic stress in the lymph nodes [3]. Some unstudied proteins, like GadB in S. dysenteriae could in this way be related to the stress response pathways [5]. The in vivo proteome reveals also very clearly the precise metabolic adaption of the pathogen [6,7]. Shigella for examples switches to an anaerobic energy metabolism inside the host and activates numerous glycolytic enzymes [5]. Similarly, the transport of various ions - iron, phosphate, copper - seems to be a major concern to the invading bacteria, proving that the adjustment of cation levels is critical for their survival. Metal-cation transporting ATPases, efflux pumps and other transporters are a common finding of several studies and M. tuberculosis’ involvement with copper might serve as a future drug target [7]. Adhesion to host cells and fellow bacteria represents another central occupation of the exploring pathogen. In N. meningitidis, different functions, such as adhesion to epithelial cells, auto-aggregation, reshaping of the host cell membrane or DNA exchange, require different, precise number of pili per bacterium. Modest decreases in pili-related proteins and thus in the “piliation index” elicit strong phenotypes, proving that determining exact protein levels can sometimes be an important clue to function [9]. Finally, an increase of ribosomal proteins and components of the translation machinery probably hint replication stages [5,6]. Interestingly, in N. meningitidis, whose main evolutionary driving force is the incorporation of exogenous DNA and homologous recombination, genes linked to the previously described functional classes, lile pili formation and adhesion, represent recombination hotspots [8]. Apart from these predictable categories, the in vivo studies unearthed some surprises, starting with a considerable number of proteins with undefined functions, which, considering the strong selection pressure exerted on pathogens, are likely to be in charge of important tasks and thus worth a closer look [4,6]. On the contrary, Shiga toxin SD1, thought to be a crucial virulence factor, turned out not to be expressed in the host [5].
ACCEPTED MANUSCRIPT
RI PT
Another important finding is that place and time matter. Liu et al. demonstrate that different locations in the same host organism induce different protein profiles [1], Kruh and colleagues have analyzed M. tuberculosis at 30 and 90 days of infection in the lungs of guinea pigs and found little overlap between the two time points. This allows many interpretations, from the emergence of drug-tolerant steady bacterial population to the drying up of the bacteria’s favorite carbon nutrients [7]. Needless to say that this adds a layer of complexity to the in vivo studies. Bacterial populations at different stages of infection probably coexist in one host organism and adapt independently to different organs and locations. The interpretation of obtained data thus request a careful spatiotemporal dissection [3,5,6].
TE D
M AN U
SC
Nevertheless, a recent study has decided to take the pathogen in vivo proteome a daring step further by looking exclusively at the surface-exposed bacterial and adherent host proteins. Rees et al. applied a technique termed “membrane shaving”, consisting in the enzymatic cleavage and subsequent identification of membraneanchored peptides, to Corynebacterium pseudotuberculosis from sheep lymph nodes [3]. The scarcity and hydrophobicity of numerous membrane proteins has hampered their detection and quantification by gel-based and MS techniques, when they are not already lost during purification, despite the fact that they are in pole position to identify unknown players of host-pathogen interactions and to provide protective antigens [4]. The study revealed about 250 bacterial and 50 host proteins. To the list of surprises adds the statement that a substantial amount of identified peptides lacks a predicted surface feature, similar to observations of bacterial exosomes, also missing known secretion signals, proving once more that the subcellular location can’t always been inferred from the genomic sequence [3,10].
AC C
EP
All in all, cell culture and in vitro studies, attempting to mimic various stress conditions, provided us with quite interesting insights into various facets of hostpathogen interactions and stress responses by the bacterium, but the big picture was still missing [6,7]. Also, due to redundancies and the complex interplay of thousands of factors, the shift to investigate a system, such as the proteome, as a whole, has greatly improved our understanding of the many faces of a bacterium [4]. In vivo analyses are the ultimate clue to the elementary physiological state of the pathogen, a glimpse at what the enemy truly looks like [6,7]. Surely, major improvements are still required. In the close future, efforts should focus on enhancing the coverage of some highly interesting subfractions of the bacterial proteome, including the “secretome”, membrane bound peptides and glycan modifications of the latter as well as pathogencontaining host cell compartments such as the phagolysosome [4,7,8]. There is a minor drawback though. As the discussed in vivo studies usually involve euthanatizing the test subject and grinding their organs, they tend to be rather animal models than humans, thus not a hundred percent representative to what will ultimately go on in the human host. Lonely planet – H. sapiens sapiens will have to wait for a while.
ACCEPTED MANUSCRIPT 1. Background
• •
Neisseria meningitidis is a commensal meningococcus colonizing the nasopharynx of healthy humans Under unclear circumstances, it can become invasive and cause septicemia or meningitis The incidence is highest in sub-Saharan Africa (1000-100 000 persons/year), affected by periodic epidemics during the dry season
RI PT
•
2. In a nutshell
• • •
SC
•
M AN U
•
Neisseria meningitidis displays distinct protein expression profiles at different infection sites in the mouse Protein abundance is high in the cerebrospinal fluid compared to blood and nasal mucosa Glutamate metabolism-associated protein expression increases in all isolates, underscoring the importance of this pathway in pathogenicity In blood, the expression of protein biogenesis components and pili proteins is enhanced PilQ favors bacterial adhesion and impedes uptake by macrophages, while GroEL is important for bacterial survival in high concentrations of human serum Deletion of NMC0101, GroEL and GuaB renders the bacterium more resistant to phagocytosis and deletion of PilQand GuaB improves survival in human serum
References
EP
Insert Cartoon nr. 26
TE D
•
AC C
[1] Liu Y, Zhang D, Engström Å, Merényi G, Hagner M, Yanh H et al. Dynamic niche-sepcific adaptations in Neisseria meningitdis during infection. Microbes Infect 2015:http://dx.doi.org/10.1016/j.micinf.2015.09.025. [2] Winnemuth M. Das große Los: Wie ich bei Günther Jauch eine halbe Million gewann und einfach losfuhr (2012). Knaus Verlag. ISBN: 978-3-8135-0504-7. [3] Rees MA, Kleifeld O, Crellin PK, Ho B, Stinear TP, Smith AI et al. Proteomic characterization of a natural host-pathogen interaction: repertoire of in vivo expressed bacterial and host surface-associated proteins. J Proteome Res 2015;14(1):120-132. [4] Schmidt F, Völker U. Proteome analysis of host-pathogen interactions: investigation of pathogen responses to the host cell environment. Proteomics 2011;11815):3203-3211.
ACCEPTED MANUSCRIPT [5] Pieper R, Zhang Q, Parmar PP, Huang ST, Clark DJ, Alami H. The Shigella dysenteriae serotype 1 proteome, profiled in the host intestinal environment, reveals major metabolic modifications and increased expression of invasive proteins. Proteomics 2009;9(22):5029-5045.
RI PT
[6] Páková, Brychta M, Strašková A, Schmidt M, Macela A, Stulík J. Comparative proteome profiling of host-pathogen interactions: insights into the adaption mechanisms of Francisella tularensis in the host cell environment. Appl Microbiol Biotechnol 2013;97(23):10103-10115. [7] Kruh NA, Troudt J, Izzo A, Prenni J, Dobos KM. Portrait of a pathogen: the Mycobacterium tuberculosis proteome in vivo. PLoS One 2010;5(11):e13938.
SC
[8] Lamelas A, Harris SR, Röltgen K, Dangy JP, Hauser J, Kigsley RA et al. Emergence of a new epidemic Neisseria meningitidis serogroup A clone in the African meningitis belt: high-resolution picture of genomic changes that mediate immune evasion. MBio 2014;5(5):e01974-019714.
M AN U
[9] Imhaus AF, Duménil G. The number of Neisseria meningitidis type IV pili determines host cell interaction. EMBO J 2014;33(16):1767-1783.
TE D
[10] Cabrita P, Trigo MJ, Ferreira RB, Brito L. Is the exoproteome important for bacterial pathogenesis? Lessons learned from interstrain exoprotein diversity in Llisteria monocytogenes grown at different temperatures. OMICS 2014;18(9):553569.
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EP
Sophia Häfner, PhD: [email protected] Postdoctoral fellow, University of Copenhagen, BRIC Biotech Research & Innovation Centre, Lund Group, 2200 Copenhagen, Denmark
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ACCEPTED MANUSCRIPT