TY  - JOUR
AU  - Mojr, Viktor
AU  - Roghanian, Mohammad
AU  - Tamman, Hedvig
AU  - Do Pham, Duy Dinh
AU  - Petrová, Magdalena
AU  - Pohl, Radek
AU  - Takada, Hiraku
AU  - Van Nerom, Katleen
AU  - Ainelo, Hanna
AU  - Caballero-Montes, Julien
AU  - Jimmy, Steffi
AU  - Garcia-Pino, Abel
AU  - Hauryliuk, Vasili
AU  - Rejman, Dominik
TI  - Nonhydrolysable Analogues of (p)ppGpp and (p)ppApp Alarmone Nucleotides as Novel Molecular Tools
JO  - ACS chemical biology
VL  - 16
IS  - 9
SN  - 1554-8929
CY  - Washington, DC
PB  - Soc.
M1  - PUBDB-2022-00522
SP  - 1680 - 1691
PY  - 2021
N1  - Waiting for fulltext
AB  - While alarmone nucleotides guanosine-3′,5′-bisdiphosphate (ppGpp) and guanosine-5′-triphosphate-3′-diphosphate (pppGpp) are archetypical bacterial second messengers, their adenosine analogues ppApp (adenosine-3′,5′-bisdiphosphate) and pppApp (adenosine-5′-triphosphate-3′-diphosphate) are toxic effectors that abrogate bacterial growth. The alarmones are both synthesized and degraded by the members of the RelA-SpoT Homologue (RSH) enzyme family. Because of the chemical and enzymatic liability of (p)ppGpp and (p)ppApp, these alarmones are prone to degradation during structural biology experiments. To overcome this limitation, we have established an efficient and straightforward procedure for synthesizing nonhydrolysable (p)ppNuNpp analogues starting from 3′-azido-3′-deoxyribonucleotides as key intermediates. To demonstrate the utility of (p)ppGNpp as a molecular tool, we show that (i) as an HD substrate mimic, ppGNpp competes with ppGpp to inhibit the enzymatic activity of human MESH1 Small Alarmone Hyrolase, SAH; and (ii) mimicking the allosteric effects of (p)ppGpp, (p)ppGNpp acts as a positive regulator of the synthetase activity of long ribosome-associated RSHs Rel and RelA. Finally, by solving the structure of the N-terminal domain region (NTD) of T. thermophilus Rel complexed with pppGNpp, we show that as an HD substrate mimic, the analogue serves as a bona fide orthosteric regulator that promotes the same intra-NTD structural rearrangements as the native substrate.
LB  - PUB:(DE-HGF)16
C6  - pmid:34477366
UR  - <Go to ISI:>//WOS:000697343400008
DO  - DOI:10.1021/acschembio.1c00398
UR  - https://bib-pubdb1.desy.de/record/474132
ER  -