Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000
Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) is a plant pathogenic bacterium that possesses complicated motility regulation pathways including a typical chemotaxis system. A significant portion of our understanding about the genes functioning in Pst DC3000 motility is based on comparison to o...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2025-01-01
|
| Series: | Frontiers in Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1535114/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832583265434730496 |
|---|---|
| author | Zichu Yang Tyler Helmann Maël Baudin Maël Baudin Maël Baudin Karl J. Schreiber Karl J. Schreiber Zhongmeng Bao Paul Stodghill Paul Stodghill Adam Deutschbauer Adam Deutschbauer Jennifer D. Lewis Jennifer D. Lewis Bryan Swingle Bryan Swingle |
| author_facet | Zichu Yang Tyler Helmann Maël Baudin Maël Baudin Maël Baudin Karl J. Schreiber Karl J. Schreiber Zhongmeng Bao Paul Stodghill Paul Stodghill Adam Deutschbauer Adam Deutschbauer Jennifer D. Lewis Jennifer D. Lewis Bryan Swingle Bryan Swingle |
| author_sort | Zichu Yang |
| collection | DOAJ |
| description | Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) is a plant pathogenic bacterium that possesses complicated motility regulation pathways including a typical chemotaxis system. A significant portion of our understanding about the genes functioning in Pst DC3000 motility is based on comparison to other bacteria. This leaves uncertainty about whether gene functions are conserved, especially since specific regulatory modules can have opposite functions in sets of Pseudomonas. In this study, we used a competitive selection to enrich for mutants with altered swimming motility and used random barcode transposon-site sequencing (RB-TnSeq) to identify genes with significant roles in swimming motility. Besides many of the known or predicted chemotaxis and motility genes, our method identified PSPTO_0406 (dipA), PSPTO_1042 (chrR) and PSPTO_4229 (hypothetical protein) as novel motility regulators. PSPTO_0406 is a homolog of dipA, a known cyclic di-GMP degrading enzyme in P. aeruginosa. PSPTO_1042 is part of an extracytoplasmic sensing system that controls gene expression in response to reactive oxygen species, suggesting that PSPTO_1042 may function as part of a mechanism that enables Pst DC3000 to alter motility when encountering oxidative stressors. PSPTO_4229 encodes a protein containing an HD-related output domain (HDOD), but with no previously identified functions. We found that deletion and overexpression of PSPTO_4229 both reduce swimming motility, suggesting that its function is sensitive to expression level. We used the overexpression phenotype to screen for nonsense and missense mutants of PSPTO_4229 that no longer reduce swimming motility and found a pair of conserved arginine residues that are necessary for motility suppression. Together these results provide a global perspective on regulatory and structural genes controlling flagellar motility in Pst DC3000. |
| format | Article |
| id | doaj-art-ea1d7c452656402a896d7dfab5da7b0e |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-ea1d7c452656402a896d7dfab5da7b0e2025-01-28T22:14:08ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011610.3389/fmicb.2025.15351141535114Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000Zichu Yang0Tyler Helmann1Maël Baudin2Maël Baudin3Maël Baudin4Karl J. Schreiber5Karl J. Schreiber6Zhongmeng Bao7Paul Stodghill8Paul Stodghill9Adam Deutschbauer10Adam Deutschbauer11Jennifer D. Lewis12Jennifer D. Lewis13Bryan Swingle14Bryan Swingle15Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, United StatesEmerging Pests and Pathogens Research Unit, Robert W. Holley Center, United States Department of Agriculture-Agricultural Research Service, Ithaca, NY, United StatesDepartment of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, United StatesPlant Gene Expression Center, United States Department of Agriculture-Agricultural Research Service, Berkeley, CA, United StatesInstitut Agro, INRAE, IRHS, SFR QUASAV, Université Angers, Angers, FranceDepartment of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, United StatesPlant Gene Expression Center, United States Department of Agriculture-Agricultural Research Service, Berkeley, CA, United StatesEmerging Pests and Pathogens Research Unit, Robert W. Holley Center, United States Department of Agriculture-Agricultural Research Service, Ithaca, NY, United StatesPlant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, United StatesEmerging Pests and Pathogens Research Unit, Robert W. Holley Center, United States Department of Agriculture-Agricultural Research Service, Ithaca, NY, United StatesDepartment of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, United StatesEnvironmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United StatesDepartment of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, United StatesPlant Gene Expression Center, United States Department of Agriculture-Agricultural Research Service, Berkeley, CA, United StatesPlant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, United StatesEmerging Pests and Pathogens Research Unit, Robert W. Holley Center, United States Department of Agriculture-Agricultural Research Service, Ithaca, NY, United StatesPseudomonas syringae pv. tomato DC3000 (Pst DC3000) is a plant pathogenic bacterium that possesses complicated motility regulation pathways including a typical chemotaxis system. A significant portion of our understanding about the genes functioning in Pst DC3000 motility is based on comparison to other bacteria. This leaves uncertainty about whether gene functions are conserved, especially since specific regulatory modules can have opposite functions in sets of Pseudomonas. In this study, we used a competitive selection to enrich for mutants with altered swimming motility and used random barcode transposon-site sequencing (RB-TnSeq) to identify genes with significant roles in swimming motility. Besides many of the known or predicted chemotaxis and motility genes, our method identified PSPTO_0406 (dipA), PSPTO_1042 (chrR) and PSPTO_4229 (hypothetical protein) as novel motility regulators. PSPTO_0406 is a homolog of dipA, a known cyclic di-GMP degrading enzyme in P. aeruginosa. PSPTO_1042 is part of an extracytoplasmic sensing system that controls gene expression in response to reactive oxygen species, suggesting that PSPTO_1042 may function as part of a mechanism that enables Pst DC3000 to alter motility when encountering oxidative stressors. PSPTO_4229 encodes a protein containing an HD-related output domain (HDOD), but with no previously identified functions. We found that deletion and overexpression of PSPTO_4229 both reduce swimming motility, suggesting that its function is sensitive to expression level. We used the overexpression phenotype to screen for nonsense and missense mutants of PSPTO_4229 that no longer reduce swimming motility and found a pair of conserved arginine residues that are necessary for motility suppression. Together these results provide a global perspective on regulatory and structural genes controlling flagellar motility in Pst DC3000.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1535114/fullPseudomonas syringaeflagellaswimming motilitychemotaxisTn-seqessential genes |
| spellingShingle | Zichu Yang Tyler Helmann Maël Baudin Maël Baudin Maël Baudin Karl J. Schreiber Karl J. Schreiber Zhongmeng Bao Paul Stodghill Paul Stodghill Adam Deutschbauer Adam Deutschbauer Jennifer D. Lewis Jennifer D. Lewis Bryan Swingle Bryan Swingle Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 Frontiers in Microbiology Pseudomonas syringae flagella swimming motility chemotaxis Tn-seq essential genes |
| title | Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 |
| title_full | Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 |
| title_fullStr | Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 |
| title_full_unstemmed | Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 |
| title_short | Genome-wide identification of novel flagellar motility genes in Pseudomonas syringae pv. tomato DC3000 |
| title_sort | genome wide identification of novel flagellar motility genes in pseudomonas syringae pv tomato dc3000 |
| topic | Pseudomonas syringae flagella swimming motility chemotaxis Tn-seq essential genes |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1535114/full |
| work_keys_str_mv | AT zichuyang genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT tylerhelmann genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT maelbaudin genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT maelbaudin genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT maelbaudin genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT karljschreiber genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT karljschreiber genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT zhongmengbao genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT paulstodghill genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT paulstodghill genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT adamdeutschbauer genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT adamdeutschbauer genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT jenniferdlewis genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT jenniferdlewis genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT bryanswingle genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 AT bryanswingle genomewideidentificationofnovelflagellarmotilitygenesinpseudomonassyringaepvtomatodc3000 |