Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.

Xanthomonas campestris strains have been reported to undergo programmed cell death (PCD) in a protein rich medium. Protein hydrolysates used in media such as nutrient broth comprise of casein digest with abundance of proline and glutamate. In the current study, X. campestris pv. campestris (Xcc) cel...

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Autores principales:	Surbhi Wadhawan, Satyendra Gautam, Arun Sharma
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cbc8532101ee4995a5d7e565dc0d70be
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spelling	oai:doaj.org-article:cbc8532101ee4995a5d7e565dc0d70be2021-11-18T08:21:15ZInvolvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.1932-620310.1371/journal.pone.0096423https://doaj.org/article/cbc8532101ee4995a5d7e565dc0d70be2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24788936/?tool=EBIhttps://doaj.org/toc/1932-6203Xanthomonas campestris strains have been reported to undergo programmed cell death (PCD) in a protein rich medium. Protein hydrolysates used in media such as nutrient broth comprise of casein digest with abundance of proline and glutamate. In the current study, X. campestris pv. campestris (Xcc) cells displayed PCD when grown in PCD inducing medium (PIM) containing casein tryptic digest. This PCD was also observed in PCD non-inducing carbohydrate rich medium (PNIM) fortified with either proline or proline along with glutamate. Surprisingly, no PCD was noticed in PNIM fortified with glutamate alone. Differential role of proline or glutamate in inducing PCD in Xcc cells growing in PNIM was studied. It was found that an intermediate product of this oxidation was involved in initiation of PCD. Proline oxidase also called as proline utilization A (PutA), catalyzes the two step oxidation of proline to glutamate. Interestingly, higher PutA activity was noticed in cells growing in PIM, and PCD was found to be inhibited by tetrahydro-2-furoic acid, a competitive inhibitor of this enzyme. Further, PCD was abolished in Xcc ΔputA strain generated using a pKNOCK suicide plasmid, and restored in Xcc ΔputA strain carrying functional PutA in a plasmid vector. Xanthomonas cells growing in PIM also displayed increased generation of ROS, as well as cell filamentation (a probable indication of SOS response). These filamented cells also displayed enhanced caspase-3-like activity during in situ labeling using a fluorescent tagged caspase-3 inhibitor (FITC-DEVD-FMK). The extent of PCD associated markers such as DNA damage, phosphatidylserine externalization and membrane depolarization were found to be significantly enhanced in wild type cells, but drastically reduced in Xcc ΔputA cells. These findings thus establish the role of PutA mediated proline oxidation in regulating death in stressed Xanthomonas cells.Surbhi WadhawanSatyendra GautamArun SharmaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 5, p e96423 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Surbhi Wadhawan Satyendra Gautam Arun Sharma Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
description	Xanthomonas campestris strains have been reported to undergo programmed cell death (PCD) in a protein rich medium. Protein hydrolysates used in media such as nutrient broth comprise of casein digest with abundance of proline and glutamate. In the current study, X. campestris pv. campestris (Xcc) cells displayed PCD when grown in PCD inducing medium (PIM) containing casein tryptic digest. This PCD was also observed in PCD non-inducing carbohydrate rich medium (PNIM) fortified with either proline or proline along with glutamate. Surprisingly, no PCD was noticed in PNIM fortified with glutamate alone. Differential role of proline or glutamate in inducing PCD in Xcc cells growing in PNIM was studied. It was found that an intermediate product of this oxidation was involved in initiation of PCD. Proline oxidase also called as proline utilization A (PutA), catalyzes the two step oxidation of proline to glutamate. Interestingly, higher PutA activity was noticed in cells growing in PIM, and PCD was found to be inhibited by tetrahydro-2-furoic acid, a competitive inhibitor of this enzyme. Further, PCD was abolished in Xcc ΔputA strain generated using a pKNOCK suicide plasmid, and restored in Xcc ΔputA strain carrying functional PutA in a plasmid vector. Xanthomonas cells growing in PIM also displayed increased generation of ROS, as well as cell filamentation (a probable indication of SOS response). These filamented cells also displayed enhanced caspase-3-like activity during in situ labeling using a fluorescent tagged caspase-3 inhibitor (FITC-DEVD-FMK). The extent of PCD associated markers such as DNA damage, phosphatidylserine externalization and membrane depolarization were found to be significantly enhanced in wild type cells, but drastically reduced in Xcc ΔputA cells. These findings thus establish the role of PutA mediated proline oxidation in regulating death in stressed Xanthomonas cells.
format	article
author	Surbhi Wadhawan Satyendra Gautam Arun Sharma
author_facet	Surbhi Wadhawan Satyendra Gautam Arun Sharma
author_sort	Surbhi Wadhawan
title	Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
title_short	Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
title_full	Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
title_fullStr	Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
title_full_unstemmed	Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.
title_sort	involvement of proline oxidase (puta) in programmed cell death of xanthomonas.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/cbc8532101ee4995a5d7e565dc0d70be
work_keys_str_mv	AT surbhiwadhawan involvementofprolineoxidaseputainprogrammedcelldeathofxanthomonas AT satyendragautam involvementofprolineoxidaseputainprogrammedcelldeathofxanthomonas AT arunsharma involvementofprolineoxidaseputainprogrammedcelldeathofxanthomonas
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Involvement of proline oxidase (PutA) in programmed cell death of Xanthomonas.

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