Fructose 1-Phosphate is the One and Only Physiological Effector
of the Cra (FruR) regulator of Pseudomonas putida.

J 2014

Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida.

CHAVARRIA, M.; G. DURANTE-RODRIGUEZ; T. KRELL; C. SANTIAGO; Jan BREZOVSKÝ et al.

Základní údaje

Originální název

Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida.

Autoři

CHAVARRIA, M.; G. DURANTE-RODRIGUEZ; T. KRELL; C. SANTIAGO; Jan BREZOVSKÝ; Jiří DAMBORSKÝ a V. DE LORENZO

Vydání

FEBS Open Bio, 2014, 2211-5463

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Velká Británie a Severní Irsko

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.515

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14310/14:00074207

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Cra; FruR; Pseudomonas putida; Fructose 1-phosphate; Fructose operon

Štítky

AKR, rivok

Změněno: 21. 1. 2015 15:36, Ing. Andrea Mikešková

Anotace

V originále

Fructose-1-phosphate (F1P) is the preferred effector of the catabolite repressor/activator (Cra) protein of the soil bacterium Pseudomonas putida but its ability to bind other metabolic intermediates in vivo is unclear. The Cra protein of this microorganism (CraPP) was submitted to mobility shift assays with target DNA sequences (the PfruB promoter) and candidate effectors fructose-1,6-biphosphate (FBP), glucose 6-phosphate (G6P), and fructose-6-phosphate (F6P). 1mM F1P was sufficient to release most of the Cra protein from its operators but more than 10mM of FBP or G6P was required to free the same complex. However, isothermal titration microcalorimetry failed to expose any specific interaction between CraPP and FBP or G6P. To solve this paradox, transcriptional activity of a PfruB-lacZ fusion was measured in wild-type and fruB cells growing on substrates that change the intracellular concentrations of F1P and FBP. The data indicated that PfruB activity was stimulated by fructose but not by glucose or succinate. This suggested that CraPP represses expression in vivo of the cognate fruBKA operon in a fashion dependent just on F1P, ruling out any other physiological effector. Molecular docking and dynamic simulations of the Cra-agonist interaction indicated that both metabolites can bind the repressor, but the breach in the relative affinity of CraPP for F1P vs FBP is three orders of magnitude larger than the equivalent distance in the Escherichia coli protein. This assigns the Cra protein of P. putida the sole role of transducing the presence of fructose in the medium into a variety of direct and indirect physiological responses.

Návaznosti

ED0001/01/01, projekt VaV

Název: CETOCOEN

EE2.3.30.0037, projekt VaV

Název: Zaměstnáním nejlepších mladých vědců k rozvoji mezinárodní spolupráce

GAP503/12/0572, projekt VaV

Název: Konstrukce syntetické metabolické dráhy pro degradaci důležitého environmentálního polutantu proteinovým a metabolickým inženýrstvím

Investor: Grantová agentura ČR, Konstrukce syntetické metabolické dráhy pro degradaci důležitého environmentlního polutantu proteinovým a metabolickým inženýrstvím

Citovat

CHAVARRIA, M.; G. DURANTE-RODRIGUEZ; T. KRELL; C. SANTIAGO; Jan BREZOVSKÝ; Jiří DAMBORSKÝ a V. DE LORENZO. Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida. FEBS Open Bio. 2014, roč. 4, č. 2014, s. 377-386. ISSN 2211-5463. Dostupné z: https://doi.org/10.1016/j.fob.2014.03.013.

@article{1213461,
   author = {Chavarria, M. and DuranteandRodriguez, G. and Krell, T. and Santiago, C. and Brezovský, Jan and Damborský, Jiří and de Lorenzo, V.},
   article_number = {2014},
   doi = {https://doi.org/10.1016/j.fob.2014.03.013},
   keywords = {Cra; FruR; Pseudomonas putida; Fructose 1-phosphate; Fructose operon},
   language = {eng},
   issn = {2211-5463},
   journal = {FEBS Open Bio},
   title = {Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida.},
   volume = {4},
   year = {2014}
}

TY  - JOUR
ID  - 1213461
AU  - Chavarria, M. - Durante-Rodriguez, G. - Krell, T. - Santiago, C. - Brezovský, Jan - Damborský, Jiří - de Lorenzo, V.
PY  - 2014
TI  - Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida.
JF  - FEBS Open Bio
VL  - 4
IS  - 2014
SP  - 377-386
EP  - 377-386
SN  - 22115463
KW  - Cra
KW  - FruR
KW  - Pseudomonas putida
KW  - Fructose 1-phosphate
KW  - Fructose operon
N2  - Fructose-1-phosphate (F1P) is the preferred effector of the catabolite repressor/activator (Cra) protein of the soil bacterium Pseudomonas putida but its ability to bind other metabolic intermediates in vivo is unclear. The Cra protein of this microorganism (CraPP) was submitted to mobility shift assays with target DNA sequences (the PfruB promoter) and candidate effectors fructose-1,6-biphosphate (FBP), glucose 6-phosphate (G6P), and fructose-6-phosphate (F6P). 1mM F1P was sufficient to release most of the Cra protein from its operators but more than 10mM of FBP or G6P was required to free the same complex. However, isothermal titration microcalorimetry failed to expose any specific interaction between CraPP and FBP or G6P. To solve this paradox, transcriptional activity of a PfruB-lacZ fusion was measured in wild-type and fruB cells growing on substrates that change the intracellular concentrations of F1P and FBP. The data indicated that PfruB activity was stimulated by fructose but not by glucose or succinate. This suggested that CraPP represses expression in vivo of the cognate fruBKA operon in a fashion dependent just on F1P, ruling out any other physiological effector. Molecular docking and dynamic simulations of the Cra-agonist interaction indicated that both metabolites can bind the repressor, but the breach in the relative affinity of CraPP for F1P vs FBP is three orders of magnitude larger than the equivalent distance in the Escherichia coli protein. This assigns the Cra protein of P. putida the sole role of transducing the presence of fructose in the medium into a variety of direct and indirect physiological responses.
ER  -

CHAVARRIA, M.; G. DURANTE-RODRIGUEZ; T. KRELL; C. SANTIAGO; Jan BREZOVSKÝ; Jiří DAMBORSKÝ a V. DE LORENZO. Fructose 1-Phosphate is the One and Only Physiological Effector of the Cra (FruR) regulator of Pseudomonas putida. \textit{FEBS Open Bio}. 2014, roč.~4, č.~2014, s.~377-386. ISSN~2211-5463. Dostupné z: https://doi.org/10.1016/j.fob.2014.03.013.

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