Gene regulation in Streptococcus pneumoniae - RePub - Erasmus ...

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Figure 3. Colonization model. Bacterial loads in the nasal lavage fluid of mice infected with (A) D39 wild-type, ΔglnA or ΔglnR and, (B) D39 wild-type, ΔglnP, ΔglnAP, and ΔgdhA. * indicates P < 0.05. Contribution of the GlnR-regulon to pneumonia In the pneumonia model the infection is monitored at three distinct sites, i.e., the nasopharynx, the lungs, and the blood compartment. This model allows assessment of the two ends of the spectrum of pneumococcal infection, i.e., the progression from nasopharyngeal colonization to invasive disease. GlnR-regulon and pneumococcal virulence In agreement with the colonization model, we observed that in the pneumonia model, mice infected with the glnA-mutant or the glnA-glnP double mutant had lower numbers of bacteria in the nasal lavage fluid at 36 post-infection than wild-type infected mice, while bacterial loads in mice infected with ΔglnR, ΔglnP or ΔgdhA did not differ from wild-type infected mice (data not shown). 167 167
Chapter 6 Figure 4. Pneumonia model. Bacterial loads in lungs and blood of mice infected with D39 wild-type, ΔglnA, ΔglnR, ΔglnP, ΔglnAP, or ΔgdhA: (A) and (C), homogenized lungs; (B) and (D), blood. * indicates P < 0.05. During the 36-h infection period, we observed no significant differences between bacterial loads in the lungs of mice infected with the glnR-mutant or wild-type bacteria (Fig. 4A). Moreover, the glnR-mutant was equally capable to enter the blood component after 12h and onwards (Fig. 4B). Similarly, we did not observe significant differences between the glnA-mutant and wild-type, although the bacterial loads in lungs and blood tended to be higher than for the glnR-mutant at later time points. However, this difference did not reach statistical significance (Fig. 4A). The glnP-mutant was attenuated after 36h of infection in the lungs, indicating that GlnP is required for full virulence in this model (Fig. 4C). In addition, in ΔglnP-infected mice fewer bacteria reached the systemic circulation, suggesting that GlnP plays a role in the dissemination of S. pneumoniae from the lungs to the blood stream (Fig. 4D). Bacterial counts of the glnA-glnP double mutant decreased in time, and were significantly lower from 12h post-infection onwards compared to the wild-type (Fig. 4C). Moreover, no bacteria reached the blood stream in mice infected with the glnA-glnP double mutant (Fig. 4D), again indicating a role for GlnP in transition from the lungs to the blood, with an additive effect of GlnA. The gdhA-mutant showed no difference in bacterial loads compared to wild-type in either lungs or systemic circulation (Fig. 4C and D), indicating that this gene is not required for full virulence in this model. 168 168
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Gene regulation in Streptococcus pn
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Cover image by Duve van Boggelen (w
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Promotiecommissie Promotoren: Prof.
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CHAPTER 1 Introduction 7
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Introduction Figure 1. Infections c
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Interestingly, autolysis induced by
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histidine kinase is the sensor prot
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metabolism are often required for v
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egulatory systems, aiming to elucid
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11. Cockeran, R., A. J. Theron, H.
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33. Li, S., S. J. Kelly, E. Lamani,
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53. Romero, P., R. Lopez, and E. Ga
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CHAPTER 2 Regulation of gene expres
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Introduction Streptococcus pneumoni
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Materiaals and Meethods Gene regula
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(Invitrogen), 0.2 mM aminoallyl dUT
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Isolation of pneumococcal RNA durin
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Resultss and Discuussion Gene regul
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Table 1. Genes regulated by RR09 in
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phosphofructokinase, and a fructose
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Figure 4. Expression of rlrA and rr
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correlated well with our in vitro m
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Figure 7. Bacterial loads in blood
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Acknowledgments This work was suppo
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10. Ibrahim, Y. M., A. R. Kerr, J.
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genome sequence of a virulent isola
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Gene regulation by RR09 in S. pneum
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Gene regulation by RR09 in S. pneum
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Table S4. Genes up- and downregulat
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Table S6. Genes up- and downregulat
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Chapter 3 Abstract 62 62 Previous s
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Chapter 3 between the wild-type and
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Chapter 3 S. pneumoniae TIGR4 by CS
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Chapter 3 performance liquid chroma
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Chapter 3 Results Transcriptional a
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Chapter 3 Table 3. Differentially e
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Chapter 3 Figure 2. Colonization mo
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Chapter 3 than the D39ΔpsaR-infect
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Chapter 3 effect was more severe fo
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Chapter 3 in TIGR4ΔpsaR. However,
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Chapter 3 References 1. Adrian, P.
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Chapter 3 21. Hemsley, C., E. Joyce
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Chapter 3 40. McCluskey, J., J. Hin
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CHAPTER 4 CodY of Streptococcus pne
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Introduction Bacteria encounter var
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Materials and Methods Bacterial str
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Table 2. Oligonucleotide primers us
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The CodY regulon of S. pneumoniae I
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The CodY regulon of S. pneumoniae w
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Accession numbers The microarray da
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Table 3. Differentially expressed g
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Binding of CodY to target promoters
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The CodY regulon of S. pneumoniae E
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The CodY regulon of S. pneumoniae F
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Discussion CodY has been described
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levels of adherence in D39. Using a
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References The CodY regulon of S. p
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Page 122 and 123: CHAPTER 5 Regulation of glutamine a
Page 124 and 125: Introduction Regulation of nitrogen
Page 126 and 127: Materials and Methods Nitrogen Meta
Page 128 and 129: TK136 D39 Δcps; Km R capsule-less
Page 130 and 131: gene from pORI28, was fused to the
Page 132 and 133: Construction of lacZ fusions Chromo
Page 134 and 135: Enzyme assays Cell-free extracts, u
Page 136 and 137: Reverse-transcriptase PCR RNA isola
Page 138 and 139: Table 3. Summary of transcriptome c
Page 140 and 141: effect caused by altered intracellu
Page 142 and 143: examine whether zwf is only transcr
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Page 146 and 147: H6-GlnR alone at the concentration
Page 148 and 149: Discussion In this study, we charac
Page 150 and 151: Acknowledgements TK, JB and HB are
Page 152 and 153: 11. den Hengst, C. D., S. A. van Hi
Page 154 and 155: Nitrogen Metabolism in S. pneumonia
Page 156 and 157: 52. Wray, L. V., Jr., J. M. Zalieck
Page 158 and 159: CHAPTER 6 Site-specific contributio
Page 160 and 161: Introduction GlnR-regulon and pneum
Page 162 and 163: Materials and Methods Bacterial str
Page 164 and 165: eferred to as t=0. Bacteriology res
Page 166 and 167: Results GlnR-regulon and pneumococc
Page 170 and 171: GlnR-regulon and pneumococcal virul
Page 172 and 173: Table 2. Differentially expressed g
Page 174 and 175: Discussion The ability to adequatel
Page 176 and 177: The microarray data showed that pre
Page 178 and 179: Acknowledgments WTH is supported by
Page 180 and 181: 12. Ibrahim, Y. M., A. R. Kerr, J.
Page 182 and 183: 33. Zalieckas, J. M., L. V. Wray, J
Page 184 and 185: CHAPTER 7 Pneumococcal gene regulat
Page 186 and 187: Introduction Regulation of gene exp
Page 188 and 189: Gene regulation and metabolism in S
Page 190 and 191: makes it possible for the pneumococ
Page 192 and 193: A CcpA homologue exists in the pneu
Page 194 and 195: double mutant, suggesting direct re
Page 196 and 197: References Gene regulation and meta
Page 198 and 199: 23. Kerr, A. R., P. V. Adrian, S. E
Page 200 and 201: 45. Sonenshein, A. L. 2005. CodY, a
Page 202 and 203: CHAPTER 8 Summarizing discussion 20
Page 204 and 205: that the observed virulence phenoty
Page 206 and 207: CodY was found to be required for a
Page 208 and 209: References Summarizing discussion 1
Page 210 and 211: Summarizing discussion 21. Orihuela
Page 212 and 213: Samenvatting en discussie Curriculu
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Curriculum Vitae Curriculum Vitae W
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Dankwoord Jeetje, dat het dan toch
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oekje en ik kijk uit naar de bijbeh
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