CH391L/S13/Mechanosensing

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Introduction

Mechanosensing refers to the ability of an organism to respond to changes in mechanical force on them or their environment. The mechanical stress can be in a variety of forms:

  • Hydrostatic pressure, as in the case of deep ocean environments
  • Fluid shear stress, as in the case of blood flowing through veins
  • Direct force, as in the case of body weight on a bone
  • Osmotic pressure, resulting from a difference in solute concentrations across a semi-permeable membrane


Effects of High Hydrostatic Pressure

High hydrostatic pressure (HHP) can cause a host of problems, including dissociation of multimeric proteins, shifts in reaction equilibria, loss of membrane integrity, and protein denaturation (reviewed in [1]). In some cases, changes in mechanical stress result in differential gene expression driven by mechanosensitive promoters or repressors. Genes that have increased expression might include cold- and heat-shock and other stress response proteins[2], barostable polymerases[3], or membrane proteins[4, 5]. Down- regulated genes might include nutrient transporters[6, 7]. In other cases, porin proteins which provide ion diffusion pathways are opened in response to osmotic stress across the membrane.

Mechanosensing Channels

Many organisms have mechanosensitive channels which are involved in changing metabolism more quickly than transcriptional changes allow. Changes in the shape of the membrane cause conformational changes that open the cahnnel, allowing solvent or solutes to flow across the membrane.

Non-Specific Channels One example of well-studied mechanosensitive channel protein is MscS (summarized in [8]). This protein forms a non-specific ion channel across the cell membrane in E. Coli which opens in response to osmotic changes in the environment. When the osmotic pressure across the membrane changes, the change in lipid bilayer structure causes MscS to open and allow ions and water to move across the membrane, which prevents cell lysis from osmotic pressure. Several other classes of mechanosensitive channels have been identified, which are generally classified by their size and the magnitude of pressure which elicits a response.

Specific Channels

Transcriptional Responses to Pressure

In Prokaryotes The first high pressure induced gene in bacteria was found in 1989[4] in a deep-ocean bacterium, Photobacterium profundum strain SS9. The gene encodes for OmpH, a large transmembrane protein which is involved in nutrient uptake. Later work found that the operon also contained two outer membrane proteins, OmpL[7] (induced at lower pressures ~1atm) and OmpI[5] (induced at much higher pressures ~400atm). Interestingly, all of these gene products encode for channel proteins, a subclass of which are the other main mechanosensing proteins. It has also been found that the lac promoter can be induced by extremely high pressures (~30 MPa) [9].

In addition to genes which are induced in response to pressure, many genes are repressed by HHP. The MalB operon is one example of a regulon repressed by high pressure [6]. Since there is no evidence of a repressor which acts only under high pressure, it is more likely that the transcription factors which induce genes constitutively are inactivated by the effects of pressure on protein folding.


In Eukaryotes

iGEM Connection

There is a brief mention of an idea regarding mechanosensing as an earthquake sensor in the notebook section for the 2012 Baskent University team, but it seems that they chose to pursue a different idea related to quorum sensing.

2010 MIT

In one of their projects, this team focused on engineering pressure sensitive promoters in mammalian cells. They were interested in using pressure as a way to control formation of biomaterials, with a goal of being able to engineer tissues and organs. Specifically, they created a touch pad which would form bone after a pressure change input. They first characterized fluid shear stress responsive promoters and used these to create a genetic toggle switch sensitive to pressure changes. Their touch pad contained human endothelial kidney (HEK) cells with production Bone Morphogenic Protein 2 (BMP2) under control of the pressure sensitive toggle switch, along with undifferentiated stem cells. When pressure is applied, BMP2 production turns on and causes the production of bone tissue via the differentiated stem cells.


2008 Tokyo Tech This team focused on creating a strain of E. Coli which could be used in a biologically-based touch screen device ("Coli Touch"). Their system applied the pressure-sensitive tet promoter to the expression of GFP. They created a device containing a thin film of cells with this construct, which resulted in the surface selectively fluorescing only in areas which had pressure applied. Since this promoter requires very high pressures (30 MPA according to their page), they also worked on using directed evolution to engineering a promoter which can respond at a lower pressure. In addition, they designed a genetic toggle switch would allow a user to "erase" any writing on the device by activating a heat-sensitive construct which represses GFP production. Beyond these experimental aspects of the project, they also developed a mathematical model which related the pressure input to fluorescence level in order to understand the limitations they needed to develop for an effective write/erase cycle using their "Coli touch" systems.

Future Directions

It's worth noting that extremely high pressure is gaining popularity as a way to pasteurize foods without heat treatment[1]. The high pressure takes less of a toll on the quality of the food products than heat, but can be just as effective at killing microorganisms. It will be interesting to see if baro-resistant organisms arise from this trend.

References

  1. Follonier S, Panke S, and Zinn M. Pressure to kill or pressure to boost: a review on the various effects and applications of hydrostatic pressure in bacterial biotechnology. Appl Microbiol Biotechnol. 2012 Mar;93(5):1805-15. DOI:10.1007/s00253-011-3854-6 | PubMed ID:22290643 | HubMed [biotechapplications]
    Review of biotechnological applications of high hydrostatic pressure
  2. Ambily Nath IV and Loka Bharathi PA. Diversity in transcripts and translational pattern of stress proteins in marine extremophiles. Extremophiles. 2011 Mar;15(2):129-53. DOI:10.1007/s00792-010-0348-x | PubMed ID:21210167 | HubMed [stressprots]
    Review of stress-induced proteins in marine extremophiles
  3. Nakasone K, Ikegami A, Fujii S, Kato C, and Horikoshi K. Isolation and piezoresponse of the rpoA gene encoding the RNA polymerase alpha subunit from the deep-sea piezophilic bacterium Shewanella violacea. FEMS Microbiol Lett. 2000 Dec 15;193(2):261-8. PubMed ID:11111034 | HubMed [RNApolviolacea]
    Study of high pressure adapted RNA polymerase from S. Violacea
  4. Bartlett D, Wright M, Yayanos AA, and Silverman M. Isolation of a gene regulated by hydrostatic pressure in a deep-sea bacterium. Nature. 1989 Nov 30;342(6249):572-4. DOI:10.1038/342572a0 | PubMed ID:2479840 | HubMed [OmpHdiscovery]
    First discovery of bacterial pressure-regulated gene, channel protein OmpH
  5. Chi E and Bartlett DH. Use of a reporter gene to follow high-pressure signal transduction in the deep-sea bacterium Photobacterium sp. strain SS9. J Bacteriol. 1993 Dec;175(23):7533-40. PubMed ID:8244922 | HubMed [OmpIgene]
    OmpL, OmpH, and OmpI identified as pressure regulated by mutational studies
  6. Sato T, Nakamura Y, Nakashima KK, Kato C, and Horikoshi K. High pressure represses expression of the malB operon in Escherichia coli. FEMS Microbiol Lett. 1996 Jan 1;135(1):111-6. PubMed ID:8598266 | HubMed [MalBinterval]
    High Hydrostatic Pressure interrupts expression of MalB operon
  7. Welch TJ and Bartlett DH. Isolation and characterization of the structural gene for OmpL, a pressure-regulated porin-like protein from the deep-sea bacterium Photobacterium species strain SS9. J Bacteriol. 1996 Aug;178(16):5027-31. PubMed ID:8759872 | HubMed [OmpLgene]
    Identification of low pressure induced channel protein
  8. Naismith JH and Booth IR. Bacterial mechanosensitive channels--MscS: evolution's solution to creating sensitivity in function. Annu Rev Biophys. 2012;41:157-77. DOI:10.1146/annurev-biophys-101211-113227 | PubMed ID:22404681 | HubMed [MscSpaper]
    Review about MscS function theories
  9. Kato C, Sato T, Smorawinska M, and Horikoshi K. High pressure conditions stimulate expression of chloramphenicol acetyltransferase regulated by the lac promoter in Escherichia coli. FEMS Microbiol Lett. 1994 Sep 15;122(1-2):91-6. PubMed ID:7958783 | HubMed [HHPlac]
    lac promoter is induced by very high pressures
  10. Morozkina EV, Slutskaia ES, Fedorova TV, Tugaĭ TI, Golubeva LI, and Koroleva OV. [Extremophilic microorganisms: biochemical adaptation and biotechnological application (review)]. Prikl Biokhim Mikrobiol. 2010 Jan-Feb;46(1):5-20. PubMed ID:20198911 | HubMed [biotechreview]
    Review of practical applications of extremophiles
  11. Oger PM and Jebbar M. The many ways of coping with pressure. Res Microbiol. 2010 Dec;161(10):799-809. DOI:10.1016/j.resmic.2010.09.017 | PubMed ID:21035541 | HubMed [copingHHP]
    Review about high hydrostatic pressure environments
  12. Schumann U, Edwards MD, Rasmussen T, Bartlett W, van West P, and Booth IR. YbdG in Escherichia coli is a threshold-setting mechanosensitive channel with MscM activity. Proc Natl Acad Sci U S A. 2010 Jul 13;107(28):12664-9. DOI:10.1073/pnas.1001405107 | PubMed ID:20616037 | HubMed [YbdG]
    Characterization of YbdG as MscM protein
  13. Levina N, Tötemeyer S, Stokes NR, Louis P, Jones MA, and Booth IR. Protection of Escherichia coli cells against extreme turgor by activation of MscS and MscL mechanosensitive channels: identification of genes required for MscS activity. EMBO J. 1999 Apr 1;18(7):1730-7. DOI:10.1093/emboj/18.7.1730 | PubMed ID:10202137 | HubMed [MscKS]
    Genes in E Coli predicted to code for MscS and MscK
All Medline abstracts: PubMed | HubMed