Molecular mutagenesis of ppGpp: turning a RelA activator into an inhibitor.
The alarmone nucleotide (p)ppGpp is a key regulator of bacterial metabolism, growth, stress tolerance and virulence, making (p)ppGpp-mediated signaling a promising target for development of antibacterials. Although ppGpp itself is an activator of the ribosome-associated ppGpp synthetase RelA, several ppGpp mimics have been developed as RelA inhibitors. However promising, the currently available ppGpp mimics are relatively inefficient, with IC50in the sub-mM range. In an attempt to identify a potent and specific inhibitor of RelA capable of abrogating (p)ppGpp production in live bacterial cells, we have tested a targeted nucleotide library using a biochemical test system comprised of purified Escherichia coli components. While none of the compounds fulfilled this aim, the screen has yielded several potentially useful molecular tools for biochemical and structural work.
University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
Jelena
J
Beljantseva
University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
Pavel
P
Kudrin
Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.
Steffi
S
Jimmy
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences v.v.i., Flemingovo nám. 2, 166 10 Prague 6, Czech Republic.
Marcel
M
Ehn
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences v.v.i., Flemingovo nám. 2, 166 10 Prague 6, Czech Republic.
Radek
R
Pohl
University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.|Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.
Vallo
V
Varik
Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama, Saitama 338-8570, Japan.
Yuzuru
Y
Tozawa
Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.
Victoria
V
Shingler
University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.
Tanel
T
Tenson
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences v.v.i., Flemingovo nám. 2, 166 10 Prague 6, Czech Republic.
Dominik
D
Rejman
University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia.|Department of Molecular Biology, Umeå University, Building 6K, 6L University Hospital Area, SE-901 87 Umeå, Sweden.|Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Building 6K and 6L, University Hospital Area, SE-901 87 Umeå, Sweden.
Vasili
V
Hauryliuk
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006159:Guanosine Tetraphosphate / Q000378:metabolism / Q000494:pharmacology; D008025:Ligases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D016296:Mutagenesis
D006159:Guanosine Tetraphosphate; D008025:Ligases; C028812:guanosine 3',5'-polyphosphate synthetases