Membrane association and polar localization of the Legionella pneumophila T4SS DotO ATPase mediated by two nonredundant receptors

被引:2
作者
Vijayrajratnam, Sukhithasri [1 ]
Milek, Sonja [2 ]
Maggi, Stefano [3 ,4 ]
Ashen, Kaleigh [1 ]
Ferrell, Micah [5 ]
Hasanovic, Ahmet [1 ]
Holgerson, Agnieszka [1 ]
Kannaiah, Shanmugapriya [1 ]
Singh, Manpreet [6 ,7 ]
Ghosal, Debnath [7 ]
Jensen, Grant J. [3 ,4 ]
Vogel, Joseph P. [1 ]
机构
[1] Washington Univ, Dept Mol Microbiol, St Louis, MO 63110 USA
[2] Univ Zurich, Balgrist Univ Hosp, Dept Neurourol, CH-8008 Zurich, Switzerland
[3] CALTECH, Div Biol & Biol Engn, Pasadena, CA 91125 USA
[4] Brigham Young Univ, Dept Chem & Biochem, Provo, UT 84602 USA
[5] Michigan State Univ, Dept Microbiol & Mol Genet, E Lansing, MI 48824 USA
[6] Univ Melbourne, Dept Biochem & Pharmacol, Bio21 Mol Sci & Biotechnol Inst, Melbourne, Vic 3010, Australia
[7] Univ Melbourne, Ctr CryoElectron Microscopy Membrane Prot, Bio21 Mol Sci & Biotechnol Inst, Australian Res Council ARC, Parkville, Vic 3010, Australia
基金
澳大利亚国家健康与医学研究理事会;
关键词
Legionella; T4SS; ATPase; receptor; secretion; IV SECRETION SYSTEM; LEGIONNAIRES-DISEASE; PLASMID R388; VIRB4; ATPASE; C-TERMINUS; PROTEIN; IDENTIFICATION; BIOGENESIS; BACTERIUM; COMPLEX;
D O I
10.1073/pnas.2401897121
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The Legionella pneumophila Dot/Icm type IVB secretion system (T4BSS) is a large, multisubunit complex that exports a vast array of substrates into eukaryotic host cells. DotO, a distant homolog of the T4ASS ATPase VirB4, associates with the bacterial inner membrane despite lacking hydrophobic transmembrane domains. Employing a genetic approach, we found DotO's membrane association is mediated by three inner-membrane Dot/Icm components, IcmT, and a combined DotJ-DotI complex (referred to as DotJI). Although deletion of icmT or dotJI individually does not affect DotO's membrane association, the simultaneous inactivation of all three genes results in increased amounts of soluble DotO. Nevertheless, deleting each receptor separately profoundly affects positioning of DotO, disrupting its link with the Dot/Icm complex at the bacterial poles, rendering the receptors nonredundant. Furthermore, a collection of dotO point mutants that we isolated established that DotO's N-terminal domain interacts with the membrane receptors and is involved in dimerization, whereas DotO's C-terminal ATPase domain primarily contributes to the protein's formation of oligomers. Modeling data revealed the complex interaction between DotO and its receptors is responsible for formation of DotO's unique "hexamer of dimers" configuration, which is a defining characteristic of VirB4 family members.
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页数:9
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