Suppressor analysis of the MotB(D33E) mutation to probe bacterial flagellar motor dynamics coupled with proton translocation.

Journal of Bacteriology
Yong-Suk CheTohru Minamino

Abstract

MotA and MotB form the stator of the proton-driven bacterial flagellar motor, which conducts protons and couples proton flow with motor rotation. Asp-33 of Salmonella enterica serovar Typhimurium MotB, which is a putative proton-binding site, is critical for torque generation. However, the mechanism of energy coupling remains unknown. Here, we carried out genetic and motility analysis of a slowly motile motB(D33E) mutant and its pseudorevertants. We first confirmed that the poor motility of the motB(D33E) mutant is due to neither protein instability, mislocalization, nor impaired interaction with MotA. We isolated 17 pseudorevertants and identified the suppressor mutations in the transmembrane helices TM2 and TM3 of MotA and in TM and the periplasmic domain of MotB. The stall torque produced by the motB(D33E) mutant motor was about half of the wild-type level, while those for the pseudorevertants were recovered nearly to the wild-type levels. However, the high-speed rotations of the motors under low-load conditions were still significantly impaired, suggesting that the rate of proton translocation is still severely limited at high speed. These results suggest that the second-site mutations recover a torque generation step invol...Continue Reading

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Citations

Jul 23, 2003·Lung Cancer : Journal of the International Association for the Study of Lung Cancer·Kazuhisa TakahashiYoshinosuke Fukuchi
Sep 30, 2010·Proceedings of the National Academy of Sciences of the United States of America·Shuichi NakamuraKeiichi Namba
Jun 22, 2013·Proceedings of the National Academy of Sciences of the United States of America·Chien-Jung LoRichard M Berry
Oct 5, 2010·Journal of Bacteriology·Fernando Suaste-OlmosLaura Camarena
Sep 11, 2012·Journal of Bacteriology·Victor Ramírez-CabreraLaura Camarena
Oct 18, 2011·Journal of Bacteriology·Daisuke Nakane, Makoto Miyata
Sep 4, 2012·International Journal of Molecular Sciences·Masayoshi Nishiyama, Seiji Kojima
Jun 28, 2014·Biomolecules·Yusuke V Morimoto, Tohru Minamino
Jul 10, 2012·Biophysical Journal·Masayoshi Nishiyama, Yoshiyuki Sowa
May 18, 2016·Structural Dynamics·Abraham Szőke, Janos Hajdu
Jan 24, 2015·Trends in Microbiology·Tohru Minamino, Katsumi Imada
Feb 26, 2015·MBio·Jonathan D PartridgeRasika M Harshey
Jul 18, 2019·ELife·Yunjie ChangJun Liu
Jul 25, 2019·Biomolecules·Shuichi Nakamura, Tohru Minamino
Oct 3, 2014·The Journal of General and Applied Microbiology·Yuka TakahashiMasahiro Ito
Jan 10, 2009·Journal of Molecular Biology·Shuichi NakamuraKeiichi Namba

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