Control of iron homeostasis in Corynebacterium glutamicum by a hierarchical regulatory network [Elektronische Ressource] / presented by Julia Frunzke

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Control of iron homeostasis in Corynebacterium glutamicum by a hierarchical regulatory network AdissertationsubmittedtotheMathematischNaturwissenschaftlicheFakultätoftheHeinrichHeineUniversitätDüsseldorfpresentedbyJuliaFrunzkeborninLichOctober2007 ThethesisinhandhasbeenperformedattheInstituteofBiotechnology1,ResearchCentreJülich,fromNovember2004untilOctober2007underthesupervisionofProf.Dr.M.Bott.Printed with acceptance of the MathematischNaturwissenschaftlichen Fakultät of theHeinrichHeineUniversitätDüsseldorf.Examiner: Prof.Dr.MichaelBott InstituteofBiotechnology1BiochemistryGroupResearchCentreJülichGmbHCoexaminer: Prof.Dr.RolfWagner DepartmentofBiophysicsHeinrichHeineUniversitätDüsseldorfOralexamination:November29,2007Results described in this dissertation have been published in the following originalpublications:Wennerhold, J.*, Krug, A.* and Bott, M. (2005) The AraCtype regulator RipA repressesaconitaseandotherironproteinsfromCorynebacteriumunderironlimitationandisitselfrepressedbyDtxR.J. Biol. Chem.280(49):4050040508.Wennerhold, J. andBott,M.(2006)TheDtxRregulonofCorynebacterium glutamicum. J. Bacteriol.188(8):29072918. Cramer, A., Auchter, M., Frunzke, J., Bott, M.
Publié le : lundi 1 janvier 2007
Lecture(s) : 17
Tags :
Source : DOCSERV.UNI-DUESSELDORF.DE/SERVLETS/DERIVATESERVLET/DERIVATE-6648/THESIS_JULIA%20FRUNZKE.PDF
Nombre de pages : 163
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Control of iron homeostasis in
Corynebacterium glutamicum
by a hierarchical regulatory network







Adissertationsubmittedtothe
MathematischNaturwissenschaftlicheFakultät
oftheHeinrichHeineUniversitätDüsseldorf



presentedby
JuliaFrunzke
borninLich


October2007

ThethesisinhandhasbeenperformedattheInstituteofBiotechnology1,ResearchCentre
Jülich,fromNovember2004untilOctober2007underthesupervisionofProf.Dr.M.Bott.




Printed with acceptance of the MathematischNaturwissenschaftlichen Fakultät of the
HeinrichHeineUniversitätDüsseldorf.









Examiner: Prof.Dr.MichaelBott
InstituteofBiotechnology1
BiochemistryGroup
ResearchCentreJülichGmbH

Coexaminer: Prof.Dr.RolfWagner
DepartmentofBiophysics
HeinrichHeineUniversitätDüsseldorf

Oralexamination:November29,2007
Results described in this dissertation have been published in the following original
publications:

Wennerhold, J.*, Krug, A.* and Bott, M. (2005) The AraCtype regulator RipA represses
aconitaseandotherironproteinsfromCorynebacteriumunderironlimitationandisitself
repressedbyDtxR.J. Biol. Chem.280(49):4050040508.

Wennerhold, J. andBott,M.(2006)TheDtxRregulonofCorynebacterium glutamicum.
J. Bacteriol.188(8):29072918.

Cramer, A., Auchter, M., Frunzke, J., Bott, M. and Eikmanns, B. J. (2006) RamB, the
transcriptionalregulatorofacetate metabolisminCorynebacterium glutamicumissubject
toregulationbyRamAandRamB.J Bacteriol189(3):11451149.

Frunzke, J.andBott,M.(2007) Evidencefortheinvolvementofthetwocomponentsystem
CgtSR11intheregulationofhemehomeostasisinCorynebacterium glutamicum
FEMS lettersto be submitted.

Frunzke, J.,Bramkamp,M.,Schweitzer,J.andBott,M. (2007) Genomeheterogeneityin
Corynebacterium glutamicum ATCC13032causedbytheprophageCGP3.J Bacteriolto be
submitted.

Frunzke, J. and Bott, M. (2008) Regulation of iron homeostasis in Corynebacterium
glutamicum. Book Chapter in “Corynebacteria: Genomics and molecular biology”.
(BurkovskiA.,ed.),HorizonScientificPress,Norwich,UK,in press.




Resultsoffurtherprojectsnotdiscussedinthisthesishavebeenpublishedin:

Frunzke, J.,Engels,V.,Hasenbein,S.,Gätgens,C.andBott,M.(2007)Coordinatedregulation
of gluconate catabolism and glucose uptake in Corynebacterium glutamicum by two
functionally equivalent transcriptional regulators, GntR1 and GntR2. Mol Microbiol in
press.




*Theseauthorscontributedequallytothiswork.
ContentI

Content

1 Summary 1


2 Introduction 2

2.1 Iron–essentialbutdangerous 2
2.2 Controlofgeneexpressionbyiron 3
2.2.1 Twofamiliesofglobalironregulators–FurandDtxR 3
2.2.2 DtxR,diphtheriatoxinrepressor 4
2.2.3 HemedependenttwocomponentsystemsinC. diphtheriae 6
2.3 IrondemandanduptakeofCorynebacterium glutamicum 7
2.4 IrondependentchangesingeneexpressioninC. glutamicum 9
2.5 Aimsofthiswork 11

3 Results 12

3.1 TheAraCtyperegulatorRipArepressesaconitaseandotheriron
proteinsfromCorynebacteriumunderironlimitationandisitself
repressedbyDtxR 14
3.2 TheDtxRregulonofCorynebacterium glutamicum 23
3.3 RamB,thetranscriptionalregulatorofacetatemetabolisminCoryne-
bacterium glutamicumissubjecttoregulationbyRamAandRamB 35
3.4 EvidencefortheinvolvementofthetwocomponentsystemCgtSR11in
theregulationofhemehomeostasisinCorynebacterium glutamicum 40
3.5 GenomeheterogeneityinCorynebacterium glutamicum ATCC13032
causedbytheprophageCGP3 51
3.6 RegulationofironhomeostasisinCorynebacterium glutamicum 64

4 Discussion 90

4.1 ControlofironhomeostasisinC. glutamicum 90
4.1.1 TheglobalironregulatorDtxR 91
4.1.2 RipA–regulatorofironproteins 95
4.1.3 ThetwocomponentsystemCgtSR11 98
4.1.4 TheArsRtyperegulatorCg0527–Subjectofinvestigations 101
4.2 FurtherputativemetaldependentregulatorsinC. glutamicum 102
4.3 InductionoftheCGP3prophageinC. glutamicum 104


IIContent

5 References 107


6 Appendix 114

6.1 SupplementalMaterial–RipA 114
6.2 SupplementalMaterial–DtxR 119
6.3 Coordinatedregulationofgluconatecatabolismandglucoseuptake
inCorynebacterium glutamicumbytwofunctionallyequivalent
transcriptionalregulators,GntR1andGntR2 128














AbbreviationsIII

Abbreviations
R Amp Ampicillinresistance
ATCC AmericanTypeCultureCollection
BHI(S) BrainHeartInfusion(+Sorbitol)
DNase Desoxyribonuclease
et al. et alii
RGm Gentamycinresistance
IPTG Isopropylthio Dgalactopyranosid
R Kan Kanamycinresistance
LB LuriaBertani
OD opticaldensityat600nm600
qRTPCR quantitativerealtimePCR
RBS ribosomebindingsite
RSpec Spectinomycinresistance
TBE TrisbaseBoricacidEDTA
TCA Trichloricacid
TE Trisbase–EDTA
TNI Trisbase–NaClImidazol
UTR untranslatedregion
UV ultraviolet
v/v volumepervolume
WT wildtype
w/v weightpervolume
YFP Yellowfluorescentprotein



Furtherabbreviationsnotincludedinthissectionareaccordingtointernationalstandards,as
forexamplelistedintheauthorguidelinesoftheFEBS Journal.

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