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@ARTICLE{Sundermeyer:911589,
author = {Sundermeyer, Lea and Bosco, Graziella and Gujar, Srushti
and Brocker, Melanie and Baumgart, Meike and Willbold,
Dieter and Weiergräber, Oliver H. and Bellinzoni, Marco and
Bott, Michael},
title = {{C}haracteristics of the {G}ln{H} and {G}ln{X} {S}ignal
{T}ransduction {P}roteins {C}ontrolling {P}kn{G}-{M}ediated
{P}hosphorylation of {O}dh{I} and 2-{O}xoglutarate
{D}ehydrogenase {A}ctivity in {C}orynebacterium glutamicum},
journal = {Microbiology spectrum},
volume = {10},
number = {6},
issn = {2165-0497},
address = {Birmingham, Ala.},
publisher = {ASM},
reportid = {FZJ-2022-04848},
pages = {e02677-22},
year = {2022},
abstract = {In Corynebacterium glutamicum the protein kinase PknG
phosphorylates OdhI and thereby abolishes the inhibition of
2-oxoglutarate dehydrogenase activity by unphosphorylated
OdhI. Our previous studies suggested that PknG activity is
controlled by the periplasmic binding protein GlnH and the
transmembrane protein GlnX, because ΔglnH and ΔglnX
mutants showed a growth defect on glutamine similar to that
of a ΔpknG mutant. We have now confirmed the involvement of
GlnH and GlnX in the control of OdhI phosphorylation by
analyzing the OdhI phosphorylation status and glutamate
secretion in ΔglnH and ΔglnX mutants and by characterizing
ΔglnX suppressor mutants. We provide evidence for GlnH
being a lipoprotein and show by isothermal titration
calorimetry that it binds l-aspartate and l-glutamate with
moderate to low affinity, but not l-glutamine, l-asparagine,
or 2-oxoglutarate. Based on a structural comparison with
GlnH of Mycobacterium tuberculosis, two residues critical
for the binding affinity were identified and verified. The
predicted GlnX topology with four transmembrane segments and
two periplasmic domains was confirmed by PhoA and LacZ
fusions. A structural model of GlnX suggested that, with the
exception of a poorly ordered N-terminal region, the entire
protein is composed of α-helices and small loops or
linkers, and it revealed similarities to other bacterial
transmembrane receptors. Our results suggest that the
GlnH-GlnX-PknG-OdhI-OdhA signal transduction cascade serves
to adapt the flux of 2-oxoglutarate between ammonium
assimilation via glutamate dehydrogenase and energy
generation via the tricarboxylic acid (TCA) cycle to the
availability of the amino group donors l-glutamate and
l-aspartate in the environment.IMPORTANCE Actinobacteria
comprise a large number of species playing important roles
in biotechnology and medicine, such as Corynebacterium
glutamicum, the major industrial amino acid producer, and
Mycobacterium tuberculosis, the pathogen causing
tuberculosis. Many actinobacteria use a signal transduction
process in which the phosphorylation status of OdhI
(corynebacteria) or GarA (mycobacteria) regulates the carbon
flux at the 2-oxoglutarate node. Inhibition of
2-oxoglutarate dehydrogenase by unphosphorylated OdhI shifts
the flux of 2-oxoglutarate from the TCA cycle toward
glutamate formation and, thus, ammonium assimilation.
Phosphorylation of OdhI/GarA is catalyzed by the protein
kinase PknG, whose activity was proposed to be controlled by
the periplasmic binding protein GlnH and the transmembrane
protein GlnX. In this study, we combined genetic,
biochemical, and structural modeling approaches to
characterize GlnH and GlnX of C. glutamicum and confirm
their roles in the GlnH-GlnX-PknG-OdhI-OdhA signal
transduction cascade. These findings are relevant also to
other Actinobacteria employing a similar control process.},
cin = {IBG-1 / IBI-7},
ddc = {570},
cid = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)IBI-7-20200312},
pnm = {5244 - Information Processing in Neuronal Networks
(POF4-524) / 2171 - Biological and environmental resources
for sustainable use (POF4-217)},
pid = {G:(DE-HGF)POF4-5244 / G:(DE-HGF)POF4-2171},
typ = {PUB:(DE-HGF)16},
pubmed = {36445153},
UT = {WOS:000891967400001},
doi = {10.1128/spectrum.02677-22},
url = {https://juser.fz-juelich.de/record/911589},
}