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Title: Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade

Abstract

Cyt1Aa is the one of four crystalline protoxins produced by mosquitocidal bacterium Bacillus thuringiensis israelensis (Bti) that has been shown to delay the evolution of insect resistance in the field. Limiting our understanding of Bti efficacy and the path to improved toxicity and spectrum has been ignorance of how Cyt1Aa crystallizes in vivo and of its mechanism of toxicity. Here, we use serial femtosecond crystallography to determine the Cyt1Aa protoxin structure from sub-micron-sized crystals produced in Bti. Structures determined under various pH/redox conditions illuminate the role played by previously uncharacterized disulfide-bridge and domain-swapped interfaces from crystal formation in Bti to dissolution in the larval mosquito midgut. Biochemical, toxicological and biophysical methods enable the deconvolution of key steps in the Cyt1Aa bioactivation cascade. We additionally show that the size, shape, production yield, pH sensitivity and toxicity of Cyt1Aa crystals grown in Bti can be controlled by single atom substitution.

Authors:
ORCiD logo [1];  [1]; ORCiD logo [1]; ORCiD logo [2];  [3]; ORCiD logo [2];  [1]; ORCiD logo [2];  [1]; ORCiD logo [4];  [1]; ORCiD logo [4];  [5]; ORCiD logo [1];  [6]; ORCiD logo [1];  [7];  [1];  [1];  [1] more »;  [1];  [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [4]; ORCiD logo [8];  [4];  [9]; ORCiD logo [9]; ORCiD logo [1];  [10]; ORCiD logo [1];  [5]; ORCiD logo [1];  [2]; ORCiD logo [1] « less
  1. Univ. Grenoble Alpes (France)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  4. European Synchrotron Radiation Facility (ESRF), Grenoble (France)
  5. Univ. of California, Riverside, CA (United States)
  6. Univ. of California, Riverside, CA (United States); California Baptist Univ., Riverside, CA (United States)
  7. Jacobs Univ., Bremen (Germany)
  8. Institut Laue-Langevin, Grenoble (France)
  9. Univ. of California, Los Angeles, CA (United States)
  10. Univ. of California, Irvine, CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); Agence Nationale de la Recherche; U.S. Centers for Disease Control and Prevention; National Institutes of Health (NIH); National Center for Advancing Translational Sciences
OSTI Identifier:
1617037
Alternate Identifier(s):
OSTI ID: 1631638
Grant/Contract Number:  
AC02-76SF00515; UO1 AI054778; RO1 AI045817; GM117126; 1U01CK000516; ANR-17-CE11-0018-01; ANR-10-INBS-05-02; ANR-17-EURE-0003; ANR-18-CE11-0005-02; AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 11; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Tetreau, Guillaume, Banneville, Anne-Sophie, Andreeva, Elena A., Brewster, Aaron S., Hunter, Mark S., Sierra, Raymond G., Teulon, Jean-Marie, Young, Iris D., Burke, Niamh, Grünewald, Tilman A., Beaudouin, Joël, Snigireva, Irina, Fernandez-Luna, Maria Teresa, Burt, Alister, Park, Hyun-Woo, Signor, Luca, Bafna, Jayesh A., Sadir, Rabia, Fenel, Daphna, Boeri-Erba, Elisabetta, Bacia, Maria, Zala, Ninon, Laporte, Frédéric, Després, Laurence, Weik, Martin, Boutet, Sébastien, Rosenthal, Martin, Coquelle, Nicolas, Burghammer, Manfred, Cascio, Duilio, Sawaya, Michael R., Winterhalter, Mathias, Gratton, Enrico, Gutsche, Irina, Federici, Brian, Pellequer, Jean-Luc, Sauter, Nicholas K., and Colletier, Jacques-Philippe. Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade. United States: N. p., 2020. Web. doi:10.1038/s41467-020-14894-w.
Tetreau, Guillaume, Banneville, Anne-Sophie, Andreeva, Elena A., Brewster, Aaron S., Hunter, Mark S., Sierra, Raymond G., Teulon, Jean-Marie, Young, Iris D., Burke, Niamh, Grünewald, Tilman A., Beaudouin, Joël, Snigireva, Irina, Fernandez-Luna, Maria Teresa, Burt, Alister, Park, Hyun-Woo, Signor, Luca, Bafna, Jayesh A., Sadir, Rabia, Fenel, Daphna, Boeri-Erba, Elisabetta, Bacia, Maria, Zala, Ninon, Laporte, Frédéric, Després, Laurence, Weik, Martin, Boutet, Sébastien, Rosenthal, Martin, Coquelle, Nicolas, Burghammer, Manfred, Cascio, Duilio, Sawaya, Michael R., Winterhalter, Mathias, Gratton, Enrico, Gutsche, Irina, Federici, Brian, Pellequer, Jean-Luc, Sauter, Nicholas K., & Colletier, Jacques-Philippe. Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade. United States. doi:https://doi.org/10.1038/s41467-020-14894-w
Tetreau, Guillaume, Banneville, Anne-Sophie, Andreeva, Elena A., Brewster, Aaron S., Hunter, Mark S., Sierra, Raymond G., Teulon, Jean-Marie, Young, Iris D., Burke, Niamh, Grünewald, Tilman A., Beaudouin, Joël, Snigireva, Irina, Fernandez-Luna, Maria Teresa, Burt, Alister, Park, Hyun-Woo, Signor, Luca, Bafna, Jayesh A., Sadir, Rabia, Fenel, Daphna, Boeri-Erba, Elisabetta, Bacia, Maria, Zala, Ninon, Laporte, Frédéric, Després, Laurence, Weik, Martin, Boutet, Sébastien, Rosenthal, Martin, Coquelle, Nicolas, Burghammer, Manfred, Cascio, Duilio, Sawaya, Michael R., Winterhalter, Mathias, Gratton, Enrico, Gutsche, Irina, Federici, Brian, Pellequer, Jean-Luc, Sauter, Nicholas K., and Colletier, Jacques-Philippe. Mon . "Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade". United States. doi:https://doi.org/10.1038/s41467-020-14894-w. https://www.osti.gov/servlets/purl/1617037.
@article{osti_1617037,
title = {Serial femtosecond crystallography on in vivo-grown crystals drives elucidation of mosquitocidal Cyt1Aa bioactivation cascade},
author = {Tetreau, Guillaume and Banneville, Anne-Sophie and Andreeva, Elena A. and Brewster, Aaron S. and Hunter, Mark S. and Sierra, Raymond G. and Teulon, Jean-Marie and Young, Iris D. and Burke, Niamh and Grünewald, Tilman A. and Beaudouin, Joël and Snigireva, Irina and Fernandez-Luna, Maria Teresa and Burt, Alister and Park, Hyun-Woo and Signor, Luca and Bafna, Jayesh A. and Sadir, Rabia and Fenel, Daphna and Boeri-Erba, Elisabetta and Bacia, Maria and Zala, Ninon and Laporte, Frédéric and Després, Laurence and Weik, Martin and Boutet, Sébastien and Rosenthal, Martin and Coquelle, Nicolas and Burghammer, Manfred and Cascio, Duilio and Sawaya, Michael R. and Winterhalter, Mathias and Gratton, Enrico and Gutsche, Irina and Federici, Brian and Pellequer, Jean-Luc and Sauter, Nicholas K. and Colletier, Jacques-Philippe},
abstractNote = {Cyt1Aa is the one of four crystalline protoxins produced by mosquitocidal bacterium Bacillus thuringiensis israelensis (Bti) that has been shown to delay the evolution of insect resistance in the field. Limiting our understanding of Bti efficacy and the path to improved toxicity and spectrum has been ignorance of how Cyt1Aa crystallizes in vivo and of its mechanism of toxicity. Here, we use serial femtosecond crystallography to determine the Cyt1Aa protoxin structure from sub-micron-sized crystals produced in Bti. Structures determined under various pH/redox conditions illuminate the role played by previously uncharacterized disulfide-bridge and domain-swapped interfaces from crystal formation in Bti to dissolution in the larval mosquito midgut. Biochemical, toxicological and biophysical methods enable the deconvolution of key steps in the Cyt1Aa bioactivation cascade. We additionally show that the size, shape, production yield, pH sensitivity and toxicity of Cyt1Aa crystals grown in Bti can be controlled by single atom substitution.},
doi = {10.1038/s41467-020-14894-w},
journal = {Nature Communications},
number = 1,
volume = 11,
place = {United States},
year = {2020},
month = {3}
}

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