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

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PsaR and pneumococcal virulence sp0798 spd0701 ciaR DNA-binding response regulator CiaR -1.1 sp0925 spd0817 hypothetical protein -1.4 sp1024 spd0910 glyA serine hydroxymethyltransferase -1.1 sp1249 spd1107 GMP reductase -1.0 sp1458 spd1287 trxB thioredoxin reductase -1.0 sp1543 spd1372 glyoxalase family protein -1.5 sp1638 spd1450 psaR iron-dependent transcriptional regulator, PsaR -2.7 -2.0 sp1802 spd1588 hypothetical protein -1.2 sp1804 spd1590 general stress protein 24, putative -1.3 sp1923 spd1726 ply pneumolysin -1.0 sp2022 spd1831 PTS system, cellobiose-specific IIC component, putative -1.4 sp2023 spd1832 PTS system, cellobiose-specific IIB component, putative -1.2 PsaR does not contribute to pneumococcal colonization To assess the contribution of PsaR-regulation to pneumococcal virulence, we examined the phenotypes of the two wild-type strains and their psaR mutants in three murine models of infection. In the colonization model, both wild-type D39 and TIGR4 were capable of extended colonization of the murine nasopharynx for a period of 192h. The level of colonization was fairly consistent during this period, varying between 10 4 - 10 6 CFU/ml (Fig. 2). We did not observe a clear effect of the deletion of psaR on the colonization ability of strains: comparable bacterial loads of wild-type and mutant strains were found during 192h of colonization, with bacterial loads between 2.5 x 10 4 CFU/ml to 3.2 x 10 5 CFU/ml for D39 wild-type and its isogenic psaR-mutant, and 1.3 x 10 5 CFU/ml to 5.0 x 10 5 CFU/ml for TIGR4 wild-type and its ΔpsaR derivative (Fig. 2A and B). The only exception was a small but significantly higher load (P=0.0049) of D39ΔpsaR at 24h (Fig.2B). 73 73
Chapter 3 Figure 2. Colonization model. Bacterial loads in the nasal lavage fluid of mice infected with (A) D39 wild-type and D39ΔpsaR, (B) TIGR4 wild-type and TIGR4ΔpsaR. * indicates P< 0.05. PsaR is not required for pneumococcal pneumonia In the pneumonia model, we observed a similar trend in bacterial loads of the nasopharynx as was observed in the colonization model, although the actual numbers of bacteria were lower. At 12h of infection there was a small but significantly higher nasopharyngeal load in mice infected with D39∆psaR as compared to wild-type (data not shown). For TIGR4 wild-type and ∆psaR, small but significant differences in bacterial load were observed throughout the entire experiment (data not shown). Furthermore, bacterial loads in the nasopharynx were comparable to those of the first 36h in the colonization model (data not shown). We did not observe any clear difference in the bacterial survival in the lungs between wild-type and psaR mutants (Fig. 3A). However, immediately after infection (0h) and 12h post-infection, the lung homogenates of D39ΔpsaR-infected mice had significantly higher bacterial loads than those of wild-type infected mice: 1.2 x 10 5 vs 6.3 x 10 4 CFU/ml at 0h, and 74 74
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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,
Page 24 and 25: 53. Romero, P., R. Lopez, and E. Ga
Page 26 and 27: CHAPTER 2 Regulation of gene expres
Page 28 and 29: Introduction Streptococcus pneumoni
Page 30 and 31: Materiaals and Meethods Gene regula
Page 32 and 33: (Invitrogen), 0.2 mM aminoallyl dUT
Page 34 and 35: Isolation of pneumococcal RNA durin
Page 36 and 37: Resultss and Discuussion Gene regul
Page 38 and 39: Table 1. Genes regulated by RR09 in
Page 40 and 41: phosphofructokinase, and a fructose
Page 42 and 43: Figure 4. Expression of rlrA and rr
Page 44 and 45: correlated well with our in vitro m
Page 46 and 47: Figure 7. Bacterial loads in blood
Page 48 and 49: Acknowledgments This work was suppo
Page 50 and 51: 10. Ibrahim, Y. M., A. R. Kerr, J.
Page 52 and 53: genome sequence of a virulent isola
Page 54 and 55: Gene regulation by RR09 in S. pneum
Page 56 and 57: Gene regulation by RR09 in S. pneum
Page 58 and 59: Table S4. Genes up- and downregulat
Page 60: Table S6. Genes up- and downregulat
Page 63 and 64: Chapter 3 Abstract 62 62 Previous s
Page 65 and 66: Chapter 3 between the wild-type and
Page 67 and 68: Chapter 3 S. pneumoniae TIGR4 by CS
Page 69 and 70: Chapter 3 performance liquid chroma
Page 71 and 72: Chapter 3 Results Transcriptional a
Page 73: Chapter 3 Table 3. Differentially e
Page 77 and 78: Chapter 3 than the D39ΔpsaR-infect
Page 79 and 80: Chapter 3 effect was more severe fo
Page 81 and 82: Chapter 3 in TIGR4ΔpsaR. However,
Page 83 and 84: Chapter 3 References 1. Adrian, P.
Page 85 and 86: Chapter 3 21. Hemsley, C., E. Joyce
Page 87 and 88: Chapter 3 40. McCluskey, J., J. Hin
Page 90 and 91: CHAPTER 4 CodY of Streptococcus pne
Page 92 and 93: Introduction Bacteria encounter var
Page 94 and 95: Materials and Methods Bacterial str
Page 96 and 97: Table 2. Oligonucleotide primers us
Page 98 and 99: The CodY regulon of S. pneumoniae I
Page 100 and 101: The CodY regulon of S. pneumoniae w
Page 102 and 103: Accession numbers The microarray da
Page 104 and 105: Table 3. Differentially expressed g
Page 106 and 107: Binding of CodY to target promoters
Page 108 and 109: The CodY regulon of S. pneumoniae E
Page 110 and 111: The CodY regulon of S. pneumoniae F
Page 112 and 113: Discussion CodY has been described
Page 114 and 115: levels of adherence in D39. Using a
Page 116 and 117: References The CodY regulon of S. p
Page 118 and 119: Glass. 2001. Genome of the bacteriu
Page 120 and 121: 40. Shivers, R. P., S. S. Dineen, a
Page 122 and 123: CHAPTER 5 Regulation of glutamine a
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Introduction Regulation of nitrogen
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Materials and Methods Nitrogen Meta
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TK136 D39 Δcps; Km R capsule-less
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gene from pORI28, was fused to the
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Construction of lacZ fusions Chromo
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Enzyme assays Cell-free extracts, u
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Reverse-transcriptase PCR RNA isola
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Table 3. Summary of transcriptome c
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effect caused by altered intracellu
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examine whether zwf is only transcr
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(http://www.bd.com/ds/technicalCent
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H6-GlnR alone at the concentration
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Discussion In this study, we charac
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Acknowledgements TK, JB and HB are
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11. den Hengst, C. D., S. A. van Hi
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Nitrogen Metabolism in S. pneumonia
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52. Wray, L. V., Jr., J. M. Zalieck
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CHAPTER 6 Site-specific contributio
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Introduction GlnR-regulon and pneum
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Materials and Methods Bacterial str
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eferred to as t=0. Bacteriology res
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Results GlnR-regulon and pneumococc
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Figure 3. Colonization model. Bacte
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GlnR-regulon and pneumococcal virul
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Table 2. Differentially expressed g
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Discussion The ability to adequatel
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The microarray data showed that pre
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Acknowledgments WTH is supported by
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12. Ibrahim, Y. M., A. R. Kerr, J.
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33. Zalieckas, J. M., L. V. Wray, J
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CHAPTER 7 Pneumococcal gene regulat
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Introduction Regulation of gene exp
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Gene regulation and metabolism in S
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makes it possible for the pneumococ
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A CcpA homologue exists in the pneu
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double mutant, suggesting direct re
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References Gene regulation and meta
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23. Kerr, A. R., P. V. Adrian, S. E
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45. Sonenshein, A. L. 2005. CodY, a
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CHAPTER 8 Summarizing discussion 20
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that the observed virulence phenoty
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CodY was found to be required for a
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References Summarizing discussion 1
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Summarizing discussion 21. Orihuela
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Samenvatting en discussie Curriculu
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Samenvatting en discussie kunnen ge
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Bevindingen die dit onderbouwen, we
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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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