cAMP dependent regulation of the TCA cycle by GlxR and oxidative stress response regulation by RosR in Corynebacterium glutamicum [Elektronische Ressource] / presented by Michael Bussmann

Micha

Sujets

• Biologie

Informations

• Publié par: heinrich-heine-universitat_dusseldorf
• Publié le: 01 janvier 2009
• Nombre de lectures: 13
• Langue: English
• Poids de l'ouvrage: 18 Mo

Extrait

cAMP dependent regulation of the TCA cycle by GlxR
and oxidative stress response regulation by RosR
in Corynebacterium glutamicum



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Examiner: Prof. Dr. Michael Bott
Institute of Biotechnology 1
Metabolic Regulation and Engineering Group
Research Centre Jülich GmbH


Coexaminer: Prof. Dr. Joachim Ernst
Department of Molecular Mycology
Heinrich-Heine-Universität Düsseldorf
























































Results described in this dissertation have been published in the following original publications:


Bussmann, M.*, Emer, D.*, Hasenbein, S., Degraf, S., Eikmanns, B. J., Bott, M. (2009)
Transcriptional control of the succinate dehydrogenase operon sdhCAB of Corynebacterium
glutamicum by the cAMP-dependent regulator GlxR and the LuxR-type regulator RamA.
J. Biotechnol. 143(3):173-82

Van Ooyen, J., Emer, D., Bussmann, M., Bott, M., Eikmanns, B. J., Eggeling, L. (2009) Citrate
synthase in Corynebacterium glutamicum is encoded by two gltA transcripts which are controlled
by RamA, RamB, and GlxR J. Biotechnol. submitted

Bussmann, M., Koch-Koerfges, A., Bott, M. (2009) Effects of a cyaB deletion on growth,
organic acid production and global gene expression in Corynebacterium glutamicum.
J. Biotechnol. to be submitted

Bussmann, M. and Bott M. (2009) RosR (Cg1324), a redox-sensing MarR-type transcriptional
regulator of Corynebacterium glutamicum. J. Biol. Chem. to be submitted







*Both authors contributed equally to this work.
































Content I
Content

1 Summary…………………………………………………………………………..……...1

2 Introduction…………………………………………………………………………...….2
2.1 The TCA cycle and its regulation in C. glutamicum……………………….……...2
2.1.1 TCA cycle and related pathways………………………………………….….…2
2.1.2 Regulation of the TCA cycle………………………………………………...….3
2.2 GlxR, a member of the cAMP receptor protein family………………………..…..6
2.3 cAMP in C. glutamicum and other bacteria……………………..……………..….8
2.4 The MarR-family of transcriptional regulators and regulation of
oxidative stress in C. glutamicum………………………………….……………..10
2.4.1 The MarR-family of transcriptional regulators………………….…………..…10
2.4.2 Regulation of oxidative stress in C. glutamicum……………….……………...11
2.5 Aims of this work………………………………………………….…………..…12
3 Results……………………………………………………………………….…………...13
3.1 Transcriptional control of the succinate dehydrogenase operon
sdhCAB of Corynebacterium glutamicum by the cAMP-dependent
regulator GlxR and the LuxR-type regulator RamA………………...............…..15
3.2 Citrate synthase in Corynebacterium glutamicum is encoded by
two gltA transcripts which are controlled by RamA, RamB, and GlxR…….........25
3.3 Effects of a cyaB deletion on growth, organic acid production and global gene
expression in Corynebacterium glutamicum…………………………..…………39
3.4 RosR (Cg1324), a redox-sensing MarR-type transcriptional regulator of
Corynebacterium glutamicum.……………....…………………………………...61

4 Discussion………………………………………………………………….……..……...82
4.1 TCA cycle regulation in C. glutamicum……………………………………..…………...82
4.1.1 GlxR, a transcriptional regulator of TCA cycle genes……………….…………..83
4.2. The adenylate cyclase CyaB in C. glutamicum……………………………………..……89
4.2.1 Characteristics of a cyaB deletion mutant…………………………..……………89
4.2.2 Decreased cAMP level effects regulation by GlxR……………….......…….……91
4.3 Regulation of oxidative stress response…………………………………………..…..….92
4.3.1 RosR a bifunctional transcriptional regulator in C.glutamicum……………….…93
4.3.2 RosR as redox-sensitive transcriptional regulator……………………………..…95 II Content

5 References……………………………………………………………………………….98

6 Appendix…………………………………………………………………………….…106
6.1 Supplemental Material-CyaB………………………….......................................106
6.2 Supplemental Material-RosR…………………………………………………...107



















Abbreviations III
Abbreviations

RAmp Ampicillin resistance
ATCC American Type Culture Collection
BHI(S) Brain Heart Infusion (+ Sorbitol)
DNase Desoxyribonuclease
DTT dithiothreitol
e.g. for example
et al. et alii
IPTG Isopropyl-thiol-ß-D-galactopyranosid
KanR Kanamycin resistance
LB Luria Bertani
MOPS 3-(N-morpholino)-propanesulfonic acid
OD600 optical density at 600 nm
ORF open reading frame
RBS ribosome binding site
rpm rounds per minute
qRT-PCR quantitative real-time PCR
SpecR Spectinomycin resistance
TBE Tris base - Boric acid – EDTA
TCA Trichloric acid
TE Tris base - EDTA
TNI Tris base - NaCl – Imidazol
UV ultraviolet
v/v volume per volume
w/v weight per volume
WT wild type
molar extinction coefficient



Abbreviations not included in this list are accordingly to international standards, as for example
listed in the author guidelines of the FEBS Journal.




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