Molecular Basis of Binding Between the Global Post-transcriptional Regulator CsrA and the T3SS Chaperone CesT

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Mar 24

PMID 29567971

Citations 17

Authors

Fei Ye

Fanli Yang

Ruijie Yu

Xi Lin

Jianxun Qi

Zhujun Chen

Yu Cao

Yuquan Wei

George F Gao

Guangwen Lu

Affiliations

Soon will be listed here.

Abstract

The T3SS chaperone CesT is recently shown to interact with the post-transcriptional regulator CsrA to modulate post-attachment signaling in enteropathogenic and enterohemorrhagic Escherichia coli. The molecular basis of the CesT/CsrA binding, however, remains elusive. Here, we show that CesT and CsrA both created two ligand binding sites in their homodimers, forming irregular multimeric complexes in solution. Through construction of a recombinant CsrA-dimer (Re-CsrA) that contains a single CesT binding site, the atomic binding features between CesT and CsrA are delineated via the structure of the CesT/Re-CsrA complex. In contrast to a previously reported N-terminally swapped dimer-form, CesT adopts a dimeric architecture with a swapped C-terminal helix for CsrA engagement. In CsrA, CesT binds to a surface patch that extensively overlaps with its mRNA binding site. The binding mode therefore justifies a mechanism of CsrA-modulation by CesT via competitive inhibition of the CsrA/mRNA interactions.

Citing Articles

Flagellar Assembly Factor FliW2 De-Represses Helicobacter pylori FlaA-Mediated Motility by Allosteric Obstruction of Global Regulator CsrA.

Su M, Dickins B, Kiang F, Tsai W, Chen Y, Chen J Helicobacter. 2025; 30(2):e70019.

PMID: 40079448 PMC: 11905337. DOI: 10.1111/hel.70019.

T3SS chaperone of the CesT family is required for secretion of the anti-sigma factor BtrA in .

Drzmisek J, Petrackova D, Dienstbier A, curnova I, Vecerek B Emerg Microbes Infect. 2023; 12(2):2272638.

PMID: 37850324 PMC: 10732220. DOI: 10.1080/22221751.2023.2272638.

Spatial Structure Formation by RsmE-Regulated Extracellular Secretions in Pseudomonas fluorescens Pf0-1.

Evans Jr A, Wells M, Denk J, Mazza W, Santos R, Delprince A J Bacteriol. 2022; 204(10):e0028522.

PMID: 36165622 PMC: 9578434. DOI: 10.1128/jb.00285-22.

Activation of the Type III Secretion System of Enteropathogenic Escherichia coli Leads to Remodeling of Its Membrane Composition and Function.

Zacharia A, Pal R, Katsowich N, Mannully C, Ibrahim A, Alfandary S mSystems. 2022; 7(3):e0020222.

PMID: 35477304 PMC: 9238428. DOI: 10.1128/msystems.00202-22.

Bacterial RNA chaperones and chaperone-like riboregulators: behind the scenes of RNA-mediated regulation of cellular metabolism.

Katsuya-Gaviria K, Paris G, Dendooven T, Bandyra K RNA Biol. 2022; 19(1):419-436.

PMID: 35438047 PMC: 9037510. DOI: 10.1080/15476286.2022.2048565.

References

Kaper J, Nataro J, Mobley H . Pathogenic Escherichia coli. Nat Rev Microbiol. 2004; 2(2):123-40. DOI: 10.1038/nrmicro818. View

Wong A, Pearson J, Bright M, Munera D, Robinson K, Lee S . Enteropathogenic and enterohaemorrhagic Escherichia coli: even more subversive elements. Mol Microbiol. 2011; 80(6):1420-38. DOI: 10.1111/j.1365-2958.2011.07661.x. View

Altegoer F, Rensing S, Bange G . Structural basis for the CsrA-dependent modulation of translation initiation by an ancient regulatory protein. Proc Natl Acad Sci U S A. 2016; 113(36):10168-73. PMC: 5018767. DOI: 10.1073/pnas.1602425113. View

Romeo T, Vakulskas C, Babitzke P . Post-transcriptional regulation on a global scale: form and function of Csr/Rsm systems. Environ Microbiol. 2012; 15(2):313-24. PMC: 3443267. DOI: 10.1111/j.1462-2920.2012.02794.x. View

Lilic M, Vujanac M, Stebbins C . A common structural motif in the binding of virulence factors to bacterial secretion chaperones. Mol Cell. 2006; 21(5):653-64. DOI: 10.1016/j.molcel.2006.01.026. View

Dubey A, Baker C, Romeo T, Babitzke P . RNA sequence and secondary structure participate in high-affinity CsrA-RNA interaction. RNA. 2005; 11(10):1579-87. PMC: 1370842. DOI: 10.1261/rna.2990205. View

Buttner C, Cornelis G, Heinz D, Niemann H . Crystal structure of Yersinia enterocolitica type III secretion chaperone SycT. Protein Sci. 2005; 14(8):1993-2002. PMC: 2279310. DOI: 10.1110/ps.051474605. View

STEBBINS C, Galan J . Maintenance of an unfolded polypeptide by a cognate chaperone in bacterial type III secretion. Nature. 2001; 414(6859):77-81. DOI: 10.1038/35102073. View

Mills E, Baruch K, Charpentier X, Kobi S, Rosenshine I . Real-time analysis of effector translocation by the type III secretion system of enteropathogenic Escherichia coli. Cell Host Microbe. 2008; 3(2):104-13. DOI: 10.1016/j.chom.2007.11.007. View

10.

Katsowich N, Elbaz N, Pal R, Mills E, Kobi S, Kahan T . Host cell attachment elicits posttranscriptional regulation in infecting enteropathogenic bacteria. Science. 2017; 355(6326):735-739. DOI: 10.1126/science.aah4886. View

11.

Mercante J, Suzuki K, Cheng X, Babitzke P, Romeo T . Comprehensive alanine-scanning mutagenesis of Escherichia coli CsrA defines two subdomains of critical functional importance. J Biol Chem. 2006; 281(42):31832-42. DOI: 10.1074/jbc.M606057200. View

12.

Thomas N, Deng W, Baker N, Puente J, Finlay B . Hierarchical delivery of an essential host colonization factor in enteropathogenic Escherichia coli. J Biol Chem. 2007; 282(40):29634-45. DOI: 10.1074/jbc.M706019200. View

13.

Delahay R, Shaw R, Elliott S, Kaper J, Knutton S, Frankel G . Functional analysis of the enteropathogenic Escherichia coli type III secretion system chaperone CesT identifies domains that mediate substrate interactions. Mol Microbiol. 2002; 43(1):61-73. DOI: 10.1046/j.1365-2958.2002.02740.x. View

14.

Creasey E, Delahay R, Bishop A, Shaw R, Kenny B, Knutton S . CesT is a bivalent enteropathogenic Escherichia coli chaperone required for translocation of both Tir and Map. Mol Microbiol. 2002; 47(1):209-21. DOI: 10.1046/j.1365-2958.2003.03290.x. View

15.

Wang Q, Qi J, Yuan Y, Xuan Y, Han P, Wan Y . Bat origins of MERS-CoV supported by bat coronavirus HKU4 usage of human receptor CD26. Cell Host Microbe. 2014; 16(3):328-37. PMC: 7104937. DOI: 10.1016/j.chom.2014.08.009. View

16.

Ramu T, Prasad M, Connors E, Mishra A, Thomassin J, Leblanc J . A novel C-terminal region within the multicargo type III secretion chaperone CesT contributes to effector secretion. J Bacteriol. 2012; 195(4):740-56. PMC: 3562101. DOI: 10.1128/JB.01967-12. View

17.

Lu G, Zhang N, Qi J, Li Y, Chen Z, Zheng C . Crystal structure of herpes simplex virus 2 gD bound to nectin-1 reveals a conserved mode of receptor recognition. J Virol. 2014; 88(23):13678-88. PMC: 4248990. DOI: 10.1128/JVI.01906-14. View

18.

Rife C, Schwarzenbacher R, McMullan D, Abdubek P, Ambing E, Axelrod H . Crystal structure of the global regulatory protein CsrA from Pseudomonas putida at 2.05 A resolution reveals a new fold. Proteins. 2005; 61(2):449-53. DOI: 10.1002/prot.20502. View

19.

Vujanac M, Stebbins C . Context-dependent protein folding of a virulence peptide in the bacterial and host environments: structure of an SycH-YopH chaperone-effector complex. Acta Crystallogr D Biol Crystallogr. 2013; 69(Pt 4):546-54. PMC: 3606036. DOI: 10.1107/S0907444912051086. View

20.

Thomas N, Deng W, Puente J, Frey E, Yip C, Strynadka N . CesT is a multi-effector chaperone and recruitment factor required for the efficient type III secretion of both LEE- and non-LEE-encoded effectors of enteropathogenic Escherichia coli. Mol Microbiol. 2005; 57(6):1762-79. DOI: 10.1111/j.1365-2958.2005.04802.x. View