Techniques to Measure Pilus Retraction Forces

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2011 Oct 14

PMID 21993648

Citations 9

Authors

Nicolas Biais

Dustin Higashi

Magdalene So

Benoit Ladoux

Affiliations

Soon will be listed here.

Abstract

The importance of physical forces in biology is becoming more appreciated. Neisseria gonorrhoeaehas become a paradigm for the study of physical forces in the bacterial world. Cycles of elongations and retractions of Type IV pili enables N. gonorrhoeaebacteria to exert forces on its environment, forces that play major roles in the life cycle of this pathogen. In order to better understand the role of these forces, there is a need to fully characterize them. Here, we present two different techniques, optical tweezers and Polyacrylamide MicroPillars (PoMPs), for measuring pilus retraction forces. Initially designed for N. gonorrhoeae, these assays can be readily modified to study other pilus-bearing bacteria including Neisseria meningitidis.

Citing Articles

An In Vitro Model System to Test Mechano-Microbiological Interactions Between Bacteria and Host Cells.

Santos L, Munteanu E, Biais N Methods Mol Biol. 2021; 2364:217-235.

PMID: 34542856 DOI: 10.1007/978-1-0716-1661-1_11.

The biochemical composition of the actomyosin network sets the magnitude of cellular traction forces.

Kollimada S, Senger F, Vignaud T, Thery M, Blanchoin L, Kurzawa L Mol Biol Cell. 2021; 32(18):1737-1748.

PMID: 34410837 PMC: 8684728. DOI: 10.1091/mbc.E21-03-0109.

Mechanomicrobiology: how bacteria sense and respond to forces.

Dufrene Y, Persat A Nat Rev Microbiol. 2020; 18(4):227-240.

PMID: 31959911 DOI: 10.1038/s41579-019-0314-2.

A bifunctional ATPase drives tad pilus extension and retraction.

Ellison C, Kan J, Chlebek J, Hummels K, Panis G, Viollier P Sci Adv. 2020; 5(12):eaay2591.

PMID: 31897429 PMC: 6920026. DOI: 10.1126/sciadv.aay2591.

PilT and PilU are homohexameric ATPases that coordinate to retract type IVa pili.

Chlebek J, Hughes H, Ratkiewicz A, Rayyan R, Wang J, Herrin B PLoS Genet. 2019; 15(10):e1008448.

PMID: 31626631 PMC: 6821130. DOI: 10.1371/journal.pgen.1008448.

References

Aas F, Egge-Jacobsen W, Winther-Larsen H, Lovold C, Hitchen P, Dell A . Neisseria gonorrhoeae type IV pili undergo multisite, hierarchical modifications with phosphoethanolamine and phosphocholine requiring an enzyme structurally related to lipopolysaccharide phosphoethanolamine transferases. J Biol Chem. 2006; 281(38):27712-23. DOI: 10.1074/jbc.M604324200. View

Hagblom P, Segal E, Billyard E, So M . Intragenic recombination leads to pilus antigenic variation in Neisseria gonorrhoeae. Nature. 1985; 315(6015):156-8. DOI: 10.1038/315156a0. View

Howie H, Glogauer M, So M . The N. gonorrhoeae type IV pilus stimulates mechanosensitive pathways and cytoprotection through a pilT-dependent mechanism. PLoS Biol. 2005; 3(4):e100. PMC: 1065265. DOI: 10.1371/journal.pbio.0030100. View

Ghassemi S, Biais N, Maniura K, Wind S, Sheetz M, Hone J . Fabrication of elastomer Pillar Arrays with Modulated Stiffness for Cellular Force Measurements. J Vac Sci Technol B Microelectron Nanometer Struct Process Meas Phenom. 2011; 26(6):2549-2553. PMC: 2880518. DOI: 10.1116/1.3013424. View

Maier B, Potter L, So M, Long C, Seifert H, Sheetz M . Single pilus motor forces exceed 100 pN. Proc Natl Acad Sci U S A. 2002; 99(25):16012-7. PMC: 138556. DOI: 10.1073/pnas.242523299. View

Sterba R, Sheetz M . Basic laser tweezers. Methods Cell Biol. 1997; 55:29-41. View

Gilbert P, Havenstrite K, Magnusson K, Sacco A, Leonardi N, Kraft P . Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture. Science. 2010; 329(5995):1078-81. PMC: 2929271. DOI: 10.1126/science.1191035. View

Biais N, Ladoux B, Higashi D, So M, Sheetz M . Cooperative retraction of bundled type IV pili enables nanonewton force generation. PLoS Biol. 2008; 6(4):e87. PMC: 2292754. DOI: 10.1371/journal.pbio.0060087. View

Simmons R, Finer J, Chu S, Spudich J . Quantitative measurements of force and displacement using an optical trap. Biophys J. 1996; 70(4):1813-22. PMC: 1225151. DOI: 10.1016/S0006-3495(96)79746-1. View

10.

du Roure O, Saez A, Buguin A, Austin R, Chavrier P, Silberzan P . Force mapping in epithelial cell migration. Proc Natl Acad Sci U S A. 2005; 102(7):2390-5. PMC: 548966. DOI: 10.1073/pnas.0408482102. View

11.

Tanase M, Biais N, Sheetz M . Magnetic tweezers in cell biology. Methods Cell Biol. 2007; 83:473-93. DOI: 10.1016/S0091-679X(07)83020-2. View

12.

Clausen M, Koomey M, Maier B . Dynamics of type IV pili is controlled by switching between multiple states. Biophys J. 2009; 96(3):1169-77. PMC: 2716576. DOI: 10.1016/j.bpj.2008.10.017. View

13.

Wang Y, Pelham Jr R . Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. Methods Enzymol. 1998; 298:489-96. DOI: 10.1016/s0076-6879(98)98041-7. View

14.

Merz A, So M, Sheetz M . Pilus retraction powers bacterial twitching motility. Nature. 2000; 407(6800):98-102. DOI: 10.1038/35024105. View

15.

Gelles J, Schnapp B, Sheetz M . Tracking kinesin-driven movements with nanometre-scale precision. Nature. 1988; 331(6155):450-3. DOI: 10.1038/331450a0. View

16.

Zhang X, Halvorsen K, Zhang C, Wong W, Springer T . Mechanoenzymatic cleavage of the ultralarge vascular protein von Willebrand factor. Science. 2009; 324(5932):1330-4. PMC: 2753189. DOI: 10.1126/science.1170905. View

17.

Kandow C, Georges P, Janmey P, Beningo K . Polyacrylamide hydrogels for cell mechanics: steps toward optimization and alternative uses. Methods Cell Biol. 2007; 83:29-46. DOI: 10.1016/S0091-679X(07)83002-0. View

18.

Higashi D, Zhang G, Biais N, Myers L, Weyand N, Elliott D . Influence of type IV pilus retraction on the architecture of the Neisseria gonorrhoeae-infected cell cortex. Microbiology (Reading). 2009; 155(Pt 12):4084-4092. PMC: 2889423. DOI: 10.1099/mic.0.032656-0. View

19.

Tan J, Tien J, Pirone D, Gray D, Bhadriraju K, Chen C . Cells lying on a bed of microneedles: an approach to isolate mechanical force. Proc Natl Acad Sci U S A. 2003; 100(4):1484-9. PMC: 149857. DOI: 10.1073/pnas.0235407100. View

20.

Farge E . Mechanical induction of Twist in the Drosophila foregut/stomodeal primordium. Curr Biol. 2003; 13(16):1365-77. DOI: 10.1016/s0960-9822(03)00576-1. View