Visualizing the Invisible: Novel Approaches to Visualizing Bacterial Proteins and Host-pathogen Interactions

Overview

Journal Front Bioeng Biotechnol

Date 2024 Feb 28

PMID 38415188

Authors

Moirangthem Kiran Singh

Linda J Kenney

Affiliations

Soon will be listed here.

Abstract

Host-pathogen interactions play a critical role in infectious diseases, and understanding the underlying mechanisms is vital for developing effective therapeutic strategies. The visualization and characterization of bacterial proteins within host cells is key to unraveling the dynamics of these interactions. Various protein labeling strategies have emerged as powerful tools for studying host-pathogen interactions, enabling the tracking, localization, and functional analysis of bacterial proteins in real-time. However, the labeling and localization of secreted type III secretion system (T3SS) effectors in host cells poses technical challenges. Conventional methods disrupt effector stoichiometry and often result in non-specific staining. Bulky fluorescent protein fusions interfere with effector secretion, while other tagging systems such as 4Cys-FLaSH/Split-GFP suffer from low labeling specificity and a poor signal-to-noise ratio. Recent advances in state-of-the-art techniques have augmented the existing toolkit for monitoring the translocation and dynamics of bacterial effectors. This comprehensive review delves into the bacterial protein labeling strategies and their application in imaging host-pathogen interactions. Lastly, we explore the obstacles faced and potential pathways forward in the realm of protein labeling strategies for visualizing interactions between hosts and pathogens.

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References

English B, Hauryliuk V, Sanamrad A, Tankov S, Dekker N, Elf J . Single-molecule investigations of the stringent response machinery in living bacterial cells. Proc Natl Acad Sci U S A. 2011; 108(31):E365-73. PMC: 3150888. DOI: 10.1073/pnas.1102255108. View

Adams S, Campbell R, Gross L, Martin B, Walkup G, Yao Y . New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications. J Am Chem Soc. 2002; 124(21):6063-76. DOI: 10.1021/ja017687n. View

Young C, Britton Z, Robinson A . Recombinant protein expression and purification: a comprehensive review of affinity tags and microbial applications. Biotechnol J. 2012; 7(5):620-34. DOI: 10.1002/biot.201100155. View

Shimada T, Takada H, Yamamoto K, Ishihama A . Expanded roles of two-component response regulator OmpR in Escherichia coli: genomic SELEX search for novel regulation targets. Genes Cells. 2015; 20(11):915-31. DOI: 10.1111/gtc.12282. View

Durisic N, Laparra-Cuervo L, Sandoval-Alvarez A, Borbely J, Lakadamyali M . Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate. Nat Methods. 2014; 11(2):156-62. DOI: 10.1038/nmeth.2784. View

Folling J, Bossi M, Bock H, Medda R, Wurm C, Hein B . Fluorescence nanoscopy by ground-state depletion and single-molecule return. Nat Methods. 2008; 5(11):943-5. DOI: 10.1038/nmeth.1257. View

Braet J, Catteeuw D, Van Damme P . Recent Advancements in Tracking Bacterial Effector Protein Translocation. Microorganisms. 2022; 10(2). PMC: 8876096. DOI: 10.3390/microorganisms10020260. View

Chin J, Santoro S, Martin A, King D, Wang L, Schultz P . Addition of p-azido-L-phenylalanine to the genetic code of Escherichia coli. J Am Chem Soc. 2002; 124(31):9026-7. DOI: 10.1021/ja027007w. View

Garner E, Bernard R, Wang W, Zhuang X, Rudner D, Mitchison T . Coupled, circumferential motions of the cell wall synthesis machinery and MreB filaments in B. subtilis. Science. 2011; 333(6039):222-5. PMC: 3235694. DOI: 10.1126/science.1203285. View

10.

Szeto J, Namolovan A, Osborne S, Coombes B, Brumell J . Salmonella-containing vacuoles display centrifugal movement associated with cell-to-cell transfer in epithelial cells. Infect Immun. 2008; 77(3):996-1007. PMC: 2643626. DOI: 10.1128/IAI.01275-08. View

11.

Perkins T, Davies M, Klemm E, Rowley G, Wileman T, James K . ChIP-seq and transcriptome analysis of the OmpR regulon of Salmonella enterica serovars Typhi and Typhimurium reveals accessory genes implicated in host colonization. Mol Microbiol. 2012; 87(3):526-38. PMC: 3586657. DOI: 10.1111/mmi.12111. View

12.

Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert L . Versatile protein tagging in cells with split fluorescent protein. Nat Commun. 2016; 7:11046. PMC: 4802074. DOI: 10.1038/ncomms11046. View

13.

Cabantous S, Terwilliger T, Waldo G . Protein tagging and detection with engineered self-assembling fragments of green fluorescent protein. Nat Biotechnol. 2004; 23(1):102-7. DOI: 10.1038/nbt1044. View

14.

Halo T, Appelbaum J, Hobert E, Balkin D, Schepartz A . Selective recognition of protein tetraserine motifs with a cell-permeable, pro-fluorescent bis-boronic acid. J Am Chem Soc. 2008; 131(2):438-9. PMC: 2659559. DOI: 10.1021/ja807872s. View

15.

Plass T, Milles S, Koehler C, Szymanski J, Mueller R, Wiessler M . Amino acids for Diels-Alder reactions in living cells. Angew Chem Int Ed Engl. 2012; 51(17):4166-70. DOI: 10.1002/anie.201108231. View

16.

Radics J, Konigsmaier L, Marlovits T . Structure of a pathogenic type 3 secretion system in action. Nat Struct Mol Biol. 2013; 21(1):82-7. DOI: 10.1038/nsmb.2722. View

17.

Enninga J, Mounier J, Sansonetti P, Tran van Nhieu G . Secretion of type III effectors into host cells in real time. Nat Methods. 2005; 2(12):959-65. DOI: 10.1038/nmeth804. View

18.

Young A, Minson M, McQuate S, Palmer A . Optimized Fluorescence Complementation Platform for Visualizing Salmonella Effector Proteins Reveals Distinctly Different Intracellular Niches in Different Cell Types. ACS Infect Dis. 2017; 3(8):575-584. PMC: 5720895. DOI: 10.1021/acsinfecdis.7b00052. View

19.

Cain R, Hayward R, Koronakis V . The target cell plasma membrane is a critical interface for Salmonella cell entry effector-host interplay. Mol Microbiol. 2004; 54(4):887-904. DOI: 10.1111/j.1365-2958.2004.04336.x. View

20.

Bujny M, Ewels P, Humphrey S, Attar N, Jepson M, Cullen P . Sorting nexin-1 defines an early phase of Salmonella-containing vacuole-remodeling during Salmonella infection. J Cell Sci. 2008; 121(Pt 12):2027-36. DOI: 10.1242/jcs.018432. View