The TatC Component of the Twin-arginine Protein Translocase Functions As an Obligate Oligomer

Overview

Journal Mol Microbiol

Specialties Microbiology
Molecular Biology

Date 2015 Jun 27

PMID 26112072

Citations 18

Authors

Francois Cleon

Johann Habersetzer

Felicity Alcock

Holger Kneuper

Phillip J Stansfeld

Hajra Basit

Mark I Wallace

Ben C Berks

Tracy Palmer

Affiliations

Soon will be listed here.

Abstract

The Tat protein export system translocates folded proteins across the bacterial cytoplasmic membrane and the plant thylakoid membrane. The Tat system in Escherichia coli is composed of TatA, TatB and TatC proteins. TatB and TatC form an oligomeric, multivalent receptor complex that binds Tat substrates, while multiple protomers of TatA assemble at substrate-bound TatBC receptors to facilitate substrate transport. We have addressed whether oligomerisation of TatC is an absolute requirement for operation of the Tat pathway by screening for dominant negative alleles of tatC that inactivate Tat function in the presence of wild-type tatC. Single substitutions that confer dominant negative TatC activity were localised to the periplasmic cap region. The variant TatC proteins retained the ability to interact with TatB and with a Tat substrate but were unable to support the in vivo assembly of TatA complexes. Blue-native PAGE analysis showed that the variant TatC proteins produced smaller TatBC complexes than the wild-type TatC protein. The substitutions did not alter disulphide crosslinking to neighbouring TatC molecules from positions in the periplasmic cap but abolished a substrate-induced disulphide crosslink in transmembrane helix 5 of TatC. Our findings show that TatC functions as an obligate oligomer.

Citing Articles

Characterization of a TatA/TatB binding site on the TatC component of the twin arginine translocase.

Severi E, Bunoro Batista M, Lannoy A, Stansfeld P, Palmer T Microbiology (Reading). 2023; 169(2).

PMID: 36790402 PMC: 10197872. DOI: 10.1099/mic.0.001298.

The temperature-dependent expression of type II secretion system controls extracellular product secretion and virulence in mesophilic SRW-OG1.

Yi X, Chen Y, Cai H, Wang J, Zhang Y, Zhu Z Front Cell Infect Microbiol. 2022; 12:945000.

PMID: 35979091 PMC: 9376225. DOI: 10.3389/fcimb.2022.945000.

A d-Phenylalanine-Benzoxazole Derivative Reveals the Role of the Essential Enzyme Rv3603c in the Pantothenate Biosynthetic Pathway of .

Pepi M, Chacko S, Marqus G, Singh V, Wang Z, Planck K ACS Infect Dis. 2022; 8(2):330-342.

PMID: 35015509 PMC: 9558617. DOI: 10.1021/acsinfecdis.1c00461.

Multicopy Suppressor Analysis of Strains Lacking Cytoplasmic Peptidyl-Prolyl Isomerases Identifies Three New PPIase Activities in That Includes the DksA Transcription Factor.

Wojtkiewicz P, Biernacka D, Gorzelak P, Stupak A, Klein G, Raina S Int J Mol Sci. 2020; 21(16).

PMID: 32823955 PMC: 7461557. DOI: 10.3390/ijms21165843.

Targeting of proteins to the twin-arginine translocation pathway.

Palmer T, Stansfeld P Mol Microbiol. 2020; 113(5):861-871.

PMID: 31971282 PMC: 7317946. DOI: 10.1111/mmi.14461.

References

Richter S, Bruser T . Targeting of unfolded PhoA to the TAT translocon of Escherichia coli. J Biol Chem. 2005; 280(52):42723-30. DOI: 10.1074/jbc.M509570200. View

Ramasamy S, Abrol R, Suloway C, Clemons Jr W . The glove-like structure of the conserved membrane protein TatC provides insight into signal sequence recognition in twin-arginine translocation. Structure. 2013; 21(5):777-88. PMC: 3653977. DOI: 10.1016/j.str.2013.03.004. View

Mori H, Cline K . A twin arginine signal peptide and the pH gradient trigger reversible assembly of the thylakoid [Delta]pH/Tat translocase. J Cell Biol. 2002; 157(2):205-10. PMC: 2199252. DOI: 10.1083/jcb.200202048. View

Oates J, Barrett C, Barnett J, Byrne K, Bolhuis A, Robinson C . The Escherichia coli twin-arginine translocation apparatus incorporates a distinct form of TatABC complex, spectrum of modular TatA complexes and minor TatAB complex. J Mol Biol. 2005; 346(1):295-305. DOI: 10.1016/j.jmb.2004.11.047. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Ma X, Cline K . Mapping the signal peptide binding and oligomer contact sites of the core subunit of the pea twin arginine protein translocase. Plant Cell. 2013; 25(3):999-1015. PMC: 3634702. DOI: 10.1105/tpc.112.107409. View

Cristobal S, de Gier J, Nielsen H, von Heijne G . Competition between Sec- and TAT-dependent protein translocation in Escherichia coli. EMBO J. 1999; 18(11):2982-90. PMC: 1171380. DOI: 10.1093/emboj/18.11.2982. View

Holzapfel E, Eisner G, Alami M, Barrett C, Buchanan G, Luke I . The entire N-terminal half of TatC is involved in twin-arginine precursor binding. Biochemistry. 2007; 46(10):2892-8. DOI: 10.1021/bi062205b. View

Deville K, Gold V, Robson A, Whitehouse S, Sessions R, Baldwin S . The oligomeric state and arrangement of the active bacterial translocon. J Biol Chem. 2010; 286(6):4659-69. PMC: 3039378. DOI: 10.1074/jbc.M110.175638. View

10.

Maldonado B, Kneuper H, Buchanan G, Hatzixanthis K, Sargent F, Berks B . Characterisation of the membrane-extrinsic domain of the TatB component of the twin arginine protein translocase. FEBS Lett. 2011; 585(3):478-84. DOI: 10.1016/j.febslet.2011.01.016. View

11.

Breyton C, Haase W, Rapoport T, Kuhlbrandt W, Collinson I . Three-dimensional structure of the bacterial protein-translocation complex SecYEG. Nature. 2002; 418(6898):662-5. DOI: 10.1038/nature00827. View

12.

Wexler M, Sargent F, Jack R, Stanley N, Bogsch E, Robinson C . TatD is a cytoplasmic protein with DNase activity. No requirement for TatD family proteins in sec-independent protein export. J Biol Chem. 2000; 275(22):16717-22. DOI: 10.1074/jbc.M000800200. View

13.

Leake M, Greene N, Godun R, Granjon T, Buchanan G, Chen S . Variable stoichiometry of the TatA component of the twin-arginine protein transport system observed by in vivo single-molecule imaging. Proc Natl Acad Sci U S A. 2008; 105(40):15376-81. PMC: 2563114. DOI: 10.1073/pnas.0806338105. View

14.

Tarry M, Schafer E, Chen S, Buchanan G, Greene N, Lea S . Structural analysis of substrate binding by the TatBC component of the twin-arginine protein transport system. Proc Natl Acad Sci U S A. 2009; 106(32):13284-9. PMC: 2718047. DOI: 10.1073/pnas.0901566106. View

15.

Dalal K, Chan C, Sligar S, Duong F . Two copies of the SecY channel and acidic lipids are necessary to activate the SecA translocation ATPase. Proc Natl Acad Sci U S A. 2012; 109(11):4104-9. PMC: 3306715. DOI: 10.1073/pnas.1117783109. View

16.

Kedrov A, Kusters I, Krasnikov V, Driessen A . A single copy of SecYEG is sufficient for preprotein translocation. EMBO J. 2011; 30(21):4387-97. PMC: 3230372. DOI: 10.1038/emboj.2011.314. View

17.

McDevitt C, Buchanan G, Sargent F, Palmer T, Berks B . Subunit composition and in vivo substrate-binding characteristics of Escherichia coli Tat protein complexes expressed at native levels. FEBS J. 2007; 273(24):5656-68. DOI: 10.1111/j.1742-4658.2006.05554.x. View

18.

Karimova G, Dautin N, Ladant D . Interaction network among Escherichia coli membrane proteins involved in cell division as revealed by bacterial two-hybrid analysis. J Bacteriol. 2005; 187(7):2233-43. PMC: 1065216. DOI: 10.1128/JB.187.7.2233-2243.2005. View

19.

Barrett C, Mangels D, Robinson C . Mutations in subunits of the Escherichia coli twin-arginine translocase block function via differing effects on translocation activity or tat complex structure. J Mol Biol. 2005; 347(2):453-63. DOI: 10.1016/j.jmb.2005.01.026. View

20.

Park E, Rapoport T . Bacterial protein translocation requires only one copy of the SecY complex in vivo. J Cell Biol. 2012; 198(5):881-93. PMC: 3432775. DOI: 10.1083/jcb.201205140. View