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

More documents

Recommendations

Info

CHAPTER 6 Site-specific contributions of glutamine-dependent regulator GlnR and GlnR-regulated genes to virulence of Streptococcus pneumoniae GlnR-regulon and pneumococcal virulence Wouter T. Hendriksen, Tomas G. Kloosterman, Hester J. Bootsma, Silvia Estevão, Ronald de Groot, Oscar P. Kuipers, and Peter W. M. Hermans. Infection and Immunity 2008, 76:1230-1238 157
Chapter 6 Abstract 158 158 The transcriptional regulator GlnR of Streptococcus pneumoniae is involved in the regulation of glutamine and glutamate metabolism, controlling the expression of the glnRA and glnPQ-zwf operons as well as the gdhA gene. To assess the contribution of the GlnR- regulon to virulence, D39 wild-type and mutants lacking genes of this regulon were tested in an in vitro adherence assay and murine infection models. All mutants, except ΔglnR, were attenuated in adherence to human pharyngeal epithelial Detroit 562 cells, suggesting contribution of these genes to adherence during colonization of humans. During murine colonization only the ΔglnA and glnP-glnA double mutant (ΔglnAP) were attenuated, in contrast to ΔglnP, indicating that the effect is caused by the lack of GlnA expression. In our pneumonia model, only ΔglnP and ΔglnAP showed a significantly reduced number of bacteria in the lungs and in blood, indicating that GlnP is required for survival in the lungs and possibly for dissemination to the blood. In intravenously infected mice, glnP and glnA are individually dispensable for survival in the blood, whereas ∆glnAP was avirulent. Finally, transcriptome analysis of the ∆glnAP showed that many genes involved in amino acid metabolism were upregulated. This signifies the importance of glutamine / glutamate uptake and synthesis for full bacterial fitness and virulence. In conclusion, several genes of the GlnR- regulon are required at different sites during pathogenesis, with glnA contributing to colonization and survival in the blood and glnP important for survival in the lungs and, possibly, efficient transition from the lungs to the blood.
Page 1 and 2:
Gene regulation in Streptococcus pn
Page 3 and 4:
Cover image by Duve van Boggelen (w
Page 5 and 6:
Promotiecommissie Promotoren: Prof.
Page 8 and 9:
CHAPTER 1 Introduction 7
Page 10 and 11:
Introduction Figure 1. Infections c
Page 12 and 13:
Interestingly, autolysis induced by
Page 14 and 15:
histidine kinase is the sensor prot
Page 16 and 17:
metabolism are often required for v
Page 18 and 19:
egulatory systems, aiming to elucid
Page 20 and 21:
11. Cockeran, R., A. J. Theron, H.
Page 22 and 23:
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 and 74:
Chapter 3 Table 3. Differentially e
Page 75 and 76:
Chapter 3 Figure 2. Colonization mo
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
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
Page 144 and 145: (http://www.bd.com/ds/technicalCent
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 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 168 and 169: Figure 3. Colonization model. Bacte
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
Page 214 and 215:
Samenvatting en discussie kunnen ge
Page 216 and 217:
Bevindingen die dit onderbouwen, we
Page 218 and 219:
Curriculum Vitae Curriculum Vitae W
Page 220 and 221:
Dankwoord Jeetje, dat het dan toch
Page 222:
oekje en ik kijk uit naar de bijbeh
show all

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

Create successful ePaper yourself

Delete template?

Save as template?