Disrupting the Acyl Carrier Protein/SpoT interaction in vivo: identification of ACP residues involved in the interaction and consequence on growth.

PloS One
Sandra AngeliniE Bouveret

Abstract

In bacteria, Acyl Carrier Protein (ACP) is the central cofactor for fatty acid biosynthesis. It carries the acyl chain in elongation and must therefore interact successively with all the enzymes of this pathway. Yet, ACP also interacts with proteins of diverse unrelated function. Among them, the interaction with SpoT has been proposed to be involved in regulating ppGpp levels in the cell in response to fatty acid synthesis inhibition. In order to better understand this mechanism, we screened for ACP mutants unable to interact with SpoT in vivo by bacterial two-hybrid, but still functional for fatty acid synthesis. The position of the selected mutations indicated that the helix II of ACP is responsible for the interaction with SpoT. This suggested a mechanism of recognition similar to one used for the enzymes of fatty acid synthesis. Consistently, the interactions tested by bacterial two-hybrid of ACP with fatty acid synthesis enzymes were also affected by the mutations that prevented the interaction with SpoT. Yet, interestingly, the corresponding mutant strains were viable, and the phenotypes of one mutant suggested a defect in growth regulation.

References

Dec 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·M SeyfzadehM Nomura
Jul 12, 1996·Biochimica Et Biophysica Acta·C O Rock, J E Cronan
May 20, 1998·Proceedings of the National Academy of Sciences of the United States of America·G KarimovaD Ladant
Jun 1, 2000·Proceedings of the National Academy of Sciences of the United States of America·K A Datsenko, B L Wanner
Oct 7, 2003·The Journal of Biological Chemistry·Yong-Mei ZhangCharles O Rock
Jul 16, 2004·Journal of Proteome Research·Mahel ZeghoufJack F Greenblatt
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Apr 28, 2005·Molecular Microbiology·Daniel VinellaRichard D'Ari
Oct 3, 2006·The Journal of Biological Chemistry·Salma RafiPeter J Tonge
May 25, 2007·The Journal of Biological Chemistry·Nicholas R De Lay, John E Cronan
Dec 7, 2007·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·David M Byers, Huansheng Gong
May 6, 2008·Annual Review of Microbiology·Katarzyna Potrykus, Michael Cashel
Sep 17, 2008·Current Protocols in Bioinformatics·Julie D ThompsonDes G Higgins
Jul 29, 2010·The Biochemical Journal·David I Chan, Hans J Vogel
Sep 21, 2010·The Journal of Biological Chemistry·Calvin A HenardAndrés Vázquez-Torres
Sep 25, 2010·The Journal of Infectious Diseases·Lee G KlinkenbergPetros C Karakousis
Oct 25, 2011·Biochemistry·Janine G BorgaroPeter J Tonge
Apr 27, 2013·Nucleic Acids Research·Undine MecholdMichael Cashel
Jul 1, 2010·World Journal of Microbiology & Biotechnology·Qi-Ming ZhouJun-Mei Zhou

❮ Previous
Next ❯

Citations

Feb 20, 2013·Trends in Microbiology·Cara C Boutte, Sean Crosson
Jul 31, 2012·Methods : a Companion to Methods in Enzymology·Aurélia Battesti, Emmanuelle Bouveret
Jul 1, 2020·PLoS Biology·María Gutiérrez-SalmerónCustodia García-Jiménez
Aug 11, 2019·Nucleic Acids Research·Karthik V RajasekarDavid J Sherratt
Dec 19, 2019·Nature Communications·Elsa GermainEtienne Maisonneuve
Oct 9, 2018·Frontiers in Microbiology·Wei ChenChristopher Davies
Nov 10, 2020·Frontiers in Microbiology·Bhabatosh Das, Rupak K Bhadra
Apr 14, 2021·Proceedings of the National Academy of Sciences of the United States of America·Terra SztainMichael D Burkart

❮ Previous
Next ❯

Methods Mentioned

BETA
tandem affinity purifications
two-hybrid
acylation
affinity co-purification
affinity purification
pull
2-hybrid
affinity
pull down
pulled down

Software Mentioned

UCSF Chimera
ClustalW
SpoT

Related Concepts

Related Feeds

Bacterial Pneumonia (ASM)

Bacterial pneumonia is a prevalent and costly infection that is a significant cause of morbidity and mortality in patients of all ages. Here is the latest research.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Bacterial Pneumonia

Bacterial pneumonia is a prevalent and costly infection that is a significant cause of morbidity and mortality in patients of all ages. Here is the latest research.

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.