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Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus
We adopted a systems-based approach to determine the role of two Candidatus Liberibacter asiaticus (CLas) proteins, LasP(235) and Effector 3, in Huanglongbing (HLB) pathogenesis. While a published work suggests the involvement of these CLas proteins HLB pathogenesis, the exact structure-based mechan...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108871/ https://www.ncbi.nlm.nih.gov/pubmed/35586217 http://dx.doi.org/10.3389/fpls.2022.869178 |
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author | Basu, Supratim Huynh, Loan Zhang, Shujian Rabara, Roel Nguyen, Hau Velásquez Guzmán, Jeanette Hao, Guixia Miles, Godfrey Shi, Qingchun Stover, Ed Gupta, Goutam |
author_facet | Basu, Supratim Huynh, Loan Zhang, Shujian Rabara, Roel Nguyen, Hau Velásquez Guzmán, Jeanette Hao, Guixia Miles, Godfrey Shi, Qingchun Stover, Ed Gupta, Goutam |
author_sort | Basu, Supratim |
collection | PubMed |
description | We adopted a systems-based approach to determine the role of two Candidatus Liberibacter asiaticus (CLas) proteins, LasP(235) and Effector 3, in Huanglongbing (HLB) pathogenesis. While a published work suggests the involvement of these CLas proteins HLB pathogenesis, the exact structure-based mechanism of their action has not been elucidated. We conducted the following experiments to determine the structure-based mechanisms of action. First, we immunoprecipitated the interacting citrus protein partners of LasP(235) and Effector 3 from the healthy and CLas-infected Hamlin extracts and identified them by Liquid Chromatography with tandem mass spectrometry (LC–MS/MS). Second, we performed a split green fluorescent protein (GFP) assay in tobacco to validate that the interactions observed in vitro are also retained in planta. The notable in planta citrus targets of LasP(235) and Effector 3 include citrus innate immune proteins. Third, in vitro and in planta studies were performed to show that LasP(235) and Effector 3 interact with and inhibit the functions of multiple citrus proteins belonging to the innate immune pathways. These inhibitory interactions led to a high level of reactive oxygen species, blocking of bactericidal lipid transfer protein (LTP), and induction of premature programed cell death (PCD), all of which are beneficial to CLas lifecycle and HLB pathogenesis. Finally, we performed molecular dynamics simulations to visualize the interactions of LasP(235) and Effector 3, respectively, with LTP and Kunitz protease inhibitor. This led to the design of an LTP mimic, which sequestered and blocked LasP(235)and rescued the bactericidal activity of LTP thereby proving that LasP(235), indeed, participates in HLB pathogenesis. |
format | Online Article Text |
id | pubmed-9108871 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-91088712022-05-17 Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus Basu, Supratim Huynh, Loan Zhang, Shujian Rabara, Roel Nguyen, Hau Velásquez Guzmán, Jeanette Hao, Guixia Miles, Godfrey Shi, Qingchun Stover, Ed Gupta, Goutam Front Plant Sci Plant Science We adopted a systems-based approach to determine the role of two Candidatus Liberibacter asiaticus (CLas) proteins, LasP(235) and Effector 3, in Huanglongbing (HLB) pathogenesis. While a published work suggests the involvement of these CLas proteins HLB pathogenesis, the exact structure-based mechanism of their action has not been elucidated. We conducted the following experiments to determine the structure-based mechanisms of action. First, we immunoprecipitated the interacting citrus protein partners of LasP(235) and Effector 3 from the healthy and CLas-infected Hamlin extracts and identified them by Liquid Chromatography with tandem mass spectrometry (LC–MS/MS). Second, we performed a split green fluorescent protein (GFP) assay in tobacco to validate that the interactions observed in vitro are also retained in planta. The notable in planta citrus targets of LasP(235) and Effector 3 include citrus innate immune proteins. Third, in vitro and in planta studies were performed to show that LasP(235) and Effector 3 interact with and inhibit the functions of multiple citrus proteins belonging to the innate immune pathways. These inhibitory interactions led to a high level of reactive oxygen species, blocking of bactericidal lipid transfer protein (LTP), and induction of premature programed cell death (PCD), all of which are beneficial to CLas lifecycle and HLB pathogenesis. Finally, we performed molecular dynamics simulations to visualize the interactions of LasP(235) and Effector 3, respectively, with LTP and Kunitz protease inhibitor. This led to the design of an LTP mimic, which sequestered and blocked LasP(235)and rescued the bactericidal activity of LTP thereby proving that LasP(235), indeed, participates in HLB pathogenesis. Frontiers Media S.A. 2022-05-02 /pmc/articles/PMC9108871/ /pubmed/35586217 http://dx.doi.org/10.3389/fpls.2022.869178 Text en Copyright © 2022 Basu, Huynh, Zhang, Rabara, Nguyen, Velásquez Guzmán, Hao, Miles, Shi, Stover and Gupta. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Basu, Supratim Huynh, Loan Zhang, Shujian Rabara, Roel Nguyen, Hau Velásquez Guzmán, Jeanette Hao, Guixia Miles, Godfrey Shi, Qingchun Stover, Ed Gupta, Goutam Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title | Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title_full | Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title_fullStr | Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title_full_unstemmed | Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title_short | Two Liberibacter Proteins Combine to Suppress Critical Innate Immune Defenses in Citrus |
title_sort | two liberibacter proteins combine to suppress critical innate immune defenses in citrus |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108871/ https://www.ncbi.nlm.nih.gov/pubmed/35586217 http://dx.doi.org/10.3389/fpls.2022.869178 |
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