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.

Basic information

Original name

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

Authors

CHAVARRIA, M. (724 Spain), G. DURANTE-RODRIGUEZ (724 Spain), T. KRELL (724 Spain), C. SANTIAGO (724 Spain), Jan BREZOVSKÝ (203 Czech Republic, belonging to the institution), Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution) and V. DE LORENZO (724 Spain)

Edition

FEBS Open Bio, 2014, 2211-5463

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 1.515

RIV identification code

RIV/00216224:14310/14:00074207

Organization unit

Faculty of Science

DOI

UT WoS

000346278200048

Keywords in English

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

Tags

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

Abstract

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.

Links

ED0001/01/01, research and development project
Name: CETOCOEN
EE2.3.30.0037, research and development project
Name: Zaměstnáním nejlepších mladých vědců k rozvoji mezinárodní spolupráce
GAP503/12/0572, research and development project
Name: Konstrukce syntetické metabolické dráhy pro degradaci důležitého environmentálního polutantu proteinovým a metabolickým inženýrstvím
Investor: Czech Science Foundation