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Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major corn pest of significant economic importance in the United States. The continuous need to control this corn maize pest and the development of field-evolved resistance toward all existing transgenic maize (Zea mays L....

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Autores principales: Kouadio, Jean-Louis, Duff, Stephen, Aikins, Michael, Zheng, Meiying, Rydel, Timothy, Chen, Danqi, Bretsnyder, Eric, Xia, Chunsheng, Zhang, Jun, Milligan, Jason, Evdokimov, Artem, Nageotte, Jeffrey, Yin, Yong, Moar, William, Giddings, Kara, Park, Yoonseong, Jerga, Agoston, Haas, Jeffrey
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8504720/
https://www.ncbi.nlm.nih.gov/pubmed/34634061
http://dx.doi.org/10.1371/journal.pone.0258052
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author Kouadio, Jean-Louis
Duff, Stephen
Aikins, Michael
Zheng, Meiying
Rydel, Timothy
Chen, Danqi
Bretsnyder, Eric
Xia, Chunsheng
Zhang, Jun
Milligan, Jason
Evdokimov, Artem
Nageotte, Jeffrey
Yin, Yong
Moar, William
Giddings, Kara
Park, Yoonseong
Jerga, Agoston
Haas, Jeffrey
author_facet Kouadio, Jean-Louis
Duff, Stephen
Aikins, Michael
Zheng, Meiying
Rydel, Timothy
Chen, Danqi
Bretsnyder, Eric
Xia, Chunsheng
Zhang, Jun
Milligan, Jason
Evdokimov, Artem
Nageotte, Jeffrey
Yin, Yong
Moar, William
Giddings, Kara
Park, Yoonseong
Jerga, Agoston
Haas, Jeffrey
author_sort Kouadio, Jean-Louis
collection PubMed
description The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major corn pest of significant economic importance in the United States. The continuous need to control this corn maize pest and the development of field-evolved resistance toward all existing transgenic maize (Zea mays L.) expressing Bacillus thuringiensis (Bt) insecticidal proteins against WCR has prompted the development of new insect-protected crops expressing distinct structural classes of insecticidal proteins. In this current study, we describe the crystal structure and functional characterization of Mpp75Aa1.1, which represents the first corn rootworm (CRW) active insecticidal protein member of the ETX_MTX2 sub-family of beta-pore forming proteins (β-PFPs), and provides new and effective protection against WCR feeding. The Mpp75Aa1.1 crystal structure was solved at 1.94 Å resolution. The Mpp75Aa1.1 is processed at its carboxyl-terminus by WCR midgut proteases, forms an oligomer, and specifically interacts with putative membrane-associated binding partners on the midgut apical microvilli to cause cellular tissue damage resulting in insect death. Alanine substitution of the surface-exposed amino acids W206, Y212, and G217 within the Mpp75Aa1.1 putative receptor binding domain I demonstrates that at least these three amino acids are required for WCR activity. The distinctive spatial arrangement of these amino acids suggests that they are part of a receptor binding epitope, which may be unique to Mpp75Aa1.1 and not present in other ETX_MTX2 proteins that do not have WCR activity. Overall, this work establishes that Mpp75Aa1.1 shares a mode of action consistent with traditional WCR-active Bt proteins despite significant structural differences.
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spelling pubmed-85047202021-10-12 Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm Kouadio, Jean-Louis Duff, Stephen Aikins, Michael Zheng, Meiying Rydel, Timothy Chen, Danqi Bretsnyder, Eric Xia, Chunsheng Zhang, Jun Milligan, Jason Evdokimov, Artem Nageotte, Jeffrey Yin, Yong Moar, William Giddings, Kara Park, Yoonseong Jerga, Agoston Haas, Jeffrey PLoS One Research Article The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is a major corn pest of significant economic importance in the United States. The continuous need to control this corn maize pest and the development of field-evolved resistance toward all existing transgenic maize (Zea mays L.) expressing Bacillus thuringiensis (Bt) insecticidal proteins against WCR has prompted the development of new insect-protected crops expressing distinct structural classes of insecticidal proteins. In this current study, we describe the crystal structure and functional characterization of Mpp75Aa1.1, which represents the first corn rootworm (CRW) active insecticidal protein member of the ETX_MTX2 sub-family of beta-pore forming proteins (β-PFPs), and provides new and effective protection against WCR feeding. The Mpp75Aa1.1 crystal structure was solved at 1.94 Å resolution. The Mpp75Aa1.1 is processed at its carboxyl-terminus by WCR midgut proteases, forms an oligomer, and specifically interacts with putative membrane-associated binding partners on the midgut apical microvilli to cause cellular tissue damage resulting in insect death. Alanine substitution of the surface-exposed amino acids W206, Y212, and G217 within the Mpp75Aa1.1 putative receptor binding domain I demonstrates that at least these three amino acids are required for WCR activity. The distinctive spatial arrangement of these amino acids suggests that they are part of a receptor binding epitope, which may be unique to Mpp75Aa1.1 and not present in other ETX_MTX2 proteins that do not have WCR activity. Overall, this work establishes that Mpp75Aa1.1 shares a mode of action consistent with traditional WCR-active Bt proteins despite significant structural differences. Public Library of Science 2021-10-11 /pmc/articles/PMC8504720/ /pubmed/34634061 http://dx.doi.org/10.1371/journal.pone.0258052 Text en © 2021 Kouadio et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kouadio, Jean-Louis
Duff, Stephen
Aikins, Michael
Zheng, Meiying
Rydel, Timothy
Chen, Danqi
Bretsnyder, Eric
Xia, Chunsheng
Zhang, Jun
Milligan, Jason
Evdokimov, Artem
Nageotte, Jeffrey
Yin, Yong
Moar, William
Giddings, Kara
Park, Yoonseong
Jerga, Agoston
Haas, Jeffrey
Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title_full Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title_fullStr Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title_full_unstemmed Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title_short Structural and functional characterization of Mpp75Aa1.1, a putative beta-pore forming protein from Brevibacillus laterosporus active against the western corn rootworm
title_sort structural and functional characterization of mpp75aa1.1, a putative beta-pore forming protein from brevibacillus laterosporus active against the western corn rootworm
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8504720/
https://www.ncbi.nlm.nih.gov/pubmed/34634061
http://dx.doi.org/10.1371/journal.pone.0258052
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