Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen
Gram-negative bacterial pathogens inject type III secreted effectors (T3SEs) directly into host cells to promote pathogen fitness by manipulating host cellular processes. Despite their crucial role in promoting virulence, relatively few T3SEs have well-characterized enzymatic activities or host targ...
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doaj-1b9e884404934edc985db608a85c9aac2021-07-02T02:37:06ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362019-02-019253554710.1534/g3.118.20087719Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic ScreenAmy Huei-Yi LeeD. Patrick BastedoJi-Young YounTimothy LoMaggie A. MiddletonInga KireevaJee Yeon LeeSara SharifpoorAnastasia BaryshnikovaJianfeng ZhangPauline W. WangSergio G. PeisajovichMichael ConstanzoBrenda J. AndrewsCharles M. BooneDarrell DesveauxDavid S. GuttmanGram-negative bacterial pathogens inject type III secreted effectors (T3SEs) directly into host cells to promote pathogen fitness by manipulating host cellular processes. Despite their crucial role in promoting virulence, relatively few T3SEs have well-characterized enzymatic activities or host targets. This is in part due to functional redundancy within pathogen T3SE repertoires as well as the promiscuity of individual T3SEs that can have multiple host targets. To overcome these challenges, we generated and characterized a collection of yeast strains stably expressing 75 T3SE constructs from the plant pathogen Pseudomonas syringae. This collection is devised to facilitate heterologous genetic screens in yeast, a non-host organism, to identify T3SEs that target conserved eukaryotic processes. Among 75 T3SEs tested, we identified 16 that inhibited yeast growth on rich media and eight that inhibited growth on stress-inducing media. We utilized Pathogenic Genetic Array (PGA) screens to identify potential host targets of P. syringae T3SEs. We focused on the acetyltransferase, HopZ1a, which interacts with plant tubulin and alters microtubule networks. To uncover putative HopZ1a host targets, we identified yeast genes with genetic interaction profiles most similar (i.e., congruent) to the PGA profile of HopZ1a and performed a functional enrichment analysis of these HopZ1a-congruent genes. We compared the congruence analyses above to previously described HopZ physical interaction datasets and identified kinesins as potential HopZ1a targets. Finally, we demonstrated that HopZ1a can target kinesins by acetylating the plant kinesins HINKEL and MKRP1, illustrating the utility of our T3SE-expressing yeast library to characterize T3SE functions.http://g3journal.org/lookup/doi/10.1534/g3.118.200877Pseudomonas syringaeType III secreted effectorPathogenic Genetic ArrayYeast screenPathogen-host interactionsHopZ1Kinesin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Amy Huei-Yi Lee D. Patrick Bastedo Ji-Young Youn Timothy Lo Maggie A. Middleton Inga Kireeva Jee Yeon Lee Sara Sharifpoor Anastasia Baryshnikova Jianfeng Zhang Pauline W. Wang Sergio G. Peisajovich Michael Constanzo Brenda J. Andrews Charles M. Boone Darrell Desveaux David S. Guttman |
spellingShingle |
Amy Huei-Yi Lee D. Patrick Bastedo Ji-Young Youn Timothy Lo Maggie A. Middleton Inga Kireeva Jee Yeon Lee Sara Sharifpoor Anastasia Baryshnikova Jianfeng Zhang Pauline W. Wang Sergio G. Peisajovich Michael Constanzo Brenda J. Andrews Charles M. Boone Darrell Desveaux David S. Guttman Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen G3: Genes, Genomes, Genetics Pseudomonas syringae Type III secreted effector Pathogenic Genetic Array Yeast screen Pathogen-host interactions HopZ1 Kinesin |
author_facet |
Amy Huei-Yi Lee D. Patrick Bastedo Ji-Young Youn Timothy Lo Maggie A. Middleton Inga Kireeva Jee Yeon Lee Sara Sharifpoor Anastasia Baryshnikova Jianfeng Zhang Pauline W. Wang Sergio G. Peisajovich Michael Constanzo Brenda J. Andrews Charles M. Boone Darrell Desveaux David S. Guttman |
author_sort |
Amy Huei-Yi Lee |
title |
Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen |
title_short |
Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen |
title_full |
Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen |
title_fullStr |
Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen |
title_full_unstemmed |
Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen |
title_sort |
identifying pseudomonas syringae type iii secreted effector function via a yeast genomic screen |
publisher |
Oxford University Press |
series |
G3: Genes, Genomes, Genetics |
issn |
2160-1836 |
publishDate |
2019-02-01 |
description |
Gram-negative bacterial pathogens inject type III secreted effectors (T3SEs) directly into host cells to promote pathogen fitness by manipulating host cellular processes. Despite their crucial role in promoting virulence, relatively few T3SEs have well-characterized enzymatic activities or host targets. This is in part due to functional redundancy within pathogen T3SE repertoires as well as the promiscuity of individual T3SEs that can have multiple host targets. To overcome these challenges, we generated and characterized a collection of yeast strains stably expressing 75 T3SE constructs from the plant pathogen Pseudomonas syringae. This collection is devised to facilitate heterologous genetic screens in yeast, a non-host organism, to identify T3SEs that target conserved eukaryotic processes. Among 75 T3SEs tested, we identified 16 that inhibited yeast growth on rich media and eight that inhibited growth on stress-inducing media. We utilized Pathogenic Genetic Array (PGA) screens to identify potential host targets of P. syringae T3SEs. We focused on the acetyltransferase, HopZ1a, which interacts with plant tubulin and alters microtubule networks. To uncover putative HopZ1a host targets, we identified yeast genes with genetic interaction profiles most similar (i.e., congruent) to the PGA profile of HopZ1a and performed a functional enrichment analysis of these HopZ1a-congruent genes. We compared the congruence analyses above to previously described HopZ physical interaction datasets and identified kinesins as potential HopZ1a targets. Finally, we demonstrated that HopZ1a can target kinesins by acetylating the plant kinesins HINKEL and MKRP1, illustrating the utility of our T3SE-expressing yeast library to characterize T3SE functions. |
topic |
Pseudomonas syringae Type III secreted effector Pathogenic Genetic Array Yeast screen Pathogen-host interactions HopZ1 Kinesin |
url |
http://g3journal.org/lookup/doi/10.1534/g3.118.200877 |
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