Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects

ABSTRACT Agrobacterium tumefaciens grows by addition of peptidoglycan (PG) at one pole of the bacterium. During the cell cycle, the cell needs to maintain two different developmental programs, one at the growth pole and another at the inert old pole. Proteins involved in this process are not yet wel...

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Autores principales:	Romain Grangeon, John Zupan, Yeonji Jeon, Patricia C. Zambryski
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Agrobacterium tumefaciens PopZ polar growth riboswitch Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c804e9c3a8de4db5818d80798d391917
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spelling	oai:doaj.org-article:c804e9c3a8de4db5818d80798d3919172021-11-15T15:51:56ZLoss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects10.1128/mBio.01881-172150-7511https://doaj.org/article/c804e9c3a8de4db5818d80798d3919172017-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01881-17https://doaj.org/toc/2150-7511ABSTRACT Agrobacterium tumefaciens grows by addition of peptidoglycan (PG) at one pole of the bacterium. During the cell cycle, the cell needs to maintain two different developmental programs, one at the growth pole and another at the inert old pole. Proteins involved in this process are not yet well characterized. To further characterize the role of pole-organizing protein A. tumefaciens PopZ (PopZAt), we created deletions of the five PopZAt domains and assayed their localization. In addition, we created a popZAt deletion strain (ΔpopZAt) that exhibited growth and cell division defects with ectopic growth poles and minicells, but the strain is unstable. To overcome the genetic instability, we created an inducible PopZAt strain by replacing the native ribosome binding site with a riboswitch. Cultivated in a medium without the inducer theophylline, the cells look like ΔpopZAt cells, with a branching and minicell phenotype. Adding theophylline restores the wild-type (WT) cell shape. Localization experiments in the depleted strain showed that the domain enriched in proline, aspartate, and glutamate likely functions in growth pole targeting. Helical domains H3 and H4 together also mediate polar localization, but only in the presence of the WT protein, suggesting that the H3 and H4 domains multimerize with WT PopZAt, to stabilize growth pole accumulation of PopZAt. IMPORTANCE Agrobacterium tumefaciens is a rod-shaped bacterium that grows by addition of PG at only one pole. The factors involved in maintaining cell asymmetry during the cell cycle with an inert old pole and a growing new pole are not well understood. Here we investigate the role of PopZAt, a homologue of Caulobacter crescentus PopZ (PopZCc), a protein essential in many aspects of pole identity in C. crescentus. We report that the loss of PopZAt leads to the appearance of branching cells, minicells, and overall growth defects. As many plant and animal pathogens also employ polar growth, understanding this process in A. tumefaciens may lead to the development of new strategies to prevent the proliferation of these pathogens. In addition, studies of A. tumefaciens will provide new insights into the evolution of the genetic networks that regulate bacterial polar growth and cell division.Romain GrangeonJohn ZupanYeonji JeonPatricia C. ZambryskiAmerican Society for MicrobiologyarticleAgrobacterium tumefaciensPopZpolar growthriboswitchMicrobiologyQR1-502ENmBio, Vol 8, Iss 6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Agrobacterium tumefaciens PopZ polar growth riboswitch Microbiology QR1-502
spellingShingle	Agrobacterium tumefaciens PopZ polar growth riboswitch Microbiology QR1-502 Romain Grangeon John Zupan Yeonji Jeon Patricia C. Zambryski Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
description	ABSTRACT Agrobacterium tumefaciens grows by addition of peptidoglycan (PG) at one pole of the bacterium. During the cell cycle, the cell needs to maintain two different developmental programs, one at the growth pole and another at the inert old pole. Proteins involved in this process are not yet well characterized. To further characterize the role of pole-organizing protein A. tumefaciens PopZ (PopZAt), we created deletions of the five PopZAt domains and assayed their localization. In addition, we created a popZAt deletion strain (ΔpopZAt) that exhibited growth and cell division defects with ectopic growth poles and minicells, but the strain is unstable. To overcome the genetic instability, we created an inducible PopZAt strain by replacing the native ribosome binding site with a riboswitch. Cultivated in a medium without the inducer theophylline, the cells look like ΔpopZAt cells, with a branching and minicell phenotype. Adding theophylline restores the wild-type (WT) cell shape. Localization experiments in the depleted strain showed that the domain enriched in proline, aspartate, and glutamate likely functions in growth pole targeting. Helical domains H3 and H4 together also mediate polar localization, but only in the presence of the WT protein, suggesting that the H3 and H4 domains multimerize with WT PopZAt, to stabilize growth pole accumulation of PopZAt. IMPORTANCE Agrobacterium tumefaciens is a rod-shaped bacterium that grows by addition of PG at only one pole. The factors involved in maintaining cell asymmetry during the cell cycle with an inert old pole and a growing new pole are not well understood. Here we investigate the role of PopZAt, a homologue of Caulobacter crescentus PopZ (PopZCc), a protein essential in many aspects of pole identity in C. crescentus. We report that the loss of PopZAt leads to the appearance of branching cells, minicells, and overall growth defects. As many plant and animal pathogens also employ polar growth, understanding this process in A. tumefaciens may lead to the development of new strategies to prevent the proliferation of these pathogens. In addition, studies of A. tumefaciens will provide new insights into the evolution of the genetic networks that regulate bacterial polar growth and cell division.
format	article
author	Romain Grangeon John Zupan Yeonji Jeon Patricia C. Zambryski
author_facet	Romain Grangeon John Zupan Yeonji Jeon Patricia C. Zambryski
author_sort	Romain Grangeon
title	Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
title_short	Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
title_full	Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
title_fullStr	Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
title_full_unstemmed	Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects
title_sort	loss of popz<sub><italic toggle="yes">at</italic></sub> activity in <italic toggle="yes">agrobacterium tumefaciens</italic> by deletion or depletion leads to multiple growth poles, minicells, and growth defects
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/c804e9c3a8de4db5818d80798d391917
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Loss of PopZ<sub><italic toggle="yes">At</italic></sub> activity in <italic toggle="yes">Agrobacterium tumefaciens</italic> by Deletion or Depletion Leads to Multiple Growth Poles, Minicells, and Growth Defects

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