Identifying Pseudomonas syringae Type III Secreted Effector Function via a Yeast Genomic Screen

G3: Genes, Genomes, Genetics

article

Amy Huei-Yi Lee¹  D. Patrick Bastedo¹  Ji-Young Youn²  Timothy Lo¹  Maggie A. Middleton³  Inga Kireeva³  Jee Yeon Lee³  Sara Sharifpoor²  Anastasia Baryshnikova⁴  Jianfeng Zhang¹  Pauline W. Wang³  Sergio G. Peisajovich¹  Michael Constanzo²  Brenda J. Andrews²  Charles M. Boone²  Darrell Desveaux¹  David S. Guttman¹ 

[1] Department of Cell & Systems Biology, University of Toronto, Toronto, Ontario, Canada;The Donnelly Centre,University of Toronto, Toronto, Ontario, Canada;Centre for the Analysis of Genome Evolution & Function, University of Toronto, Toronto, Ontario, Canada;Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey;Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada

关键词: Pseudomonas syringae;    Type III secreted effector;    Pathogenic Genetic Array;    Yeast screen;    Pathogen-host interactions;    HopZ1;    Kinesin;   

DOI  :  10.1534/g3.118.200877

学科分类：社会科学、人文和艺术（综合）

来源: Genetics Society of America

【 摘 要 】

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.

【 授权许可】

CC BY|CC BY-NC