Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium

Abstract Ehrlichia chaffeensis, a tick-transmitted rickettsial bacterium, is the causative agent of human monocytic ehrlichiosis. Biochemical characterization of this and other related Rickettsiales remains a major challenge, as they require a host cell for their replication. We investigated the use...

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Autores principales:	Vijay K. Eedunuri, Yuntao Zhang, Chuanmin Cheng, Li Chen, Huitao Liu, Anders Omsland, Dan Boyle, Roman R. Ganta
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5d6953aeb10646b4ba0e1e46b4cb5c76
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spelling	oai:doaj.org-article:5d6953aeb10646b4ba0e1e46b4cb5c762021-12-02T15:07:47ZProtein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium10.1038/s41598-018-27574-z2045-2322https://doaj.org/article/5d6953aeb10646b4ba0e1e46b4cb5c762018-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-27574-zhttps://doaj.org/toc/2045-2322Abstract Ehrlichia chaffeensis, a tick-transmitted rickettsial bacterium, is the causative agent of human monocytic ehrlichiosis. Biochemical characterization of this and other related Rickettsiales remains a major challenge, as they require a host cell for their replication. We investigated the use of an axenic medium for E. chaffeensis growth, assessed by protein and DNA synthesis, in the absence of a host cell. E. chaffeensis organisms harvested from in vitro cultures grown in a vertebrate cell line were fractionated into infectious dense-core cells (DC) and the non-infectious replicating form, known as reticulate cells (RC) by renografin density gradient centrifugation and incubated in the axenic medium containing amino acids, nucleotides, and different energy sources. Bacterial protein and DNA synthesis were observed in RCs in response to glucose-6-phosphate, although adenosine triphosphate, alpha-ketoglutarate or sodium acetate supported protein synthesis. The biosynthetic activity could not be detected in DCs in the axenic medium. While the data demonstrate de novo protein and DNA synthesis under axenic conditions for E. chaffeensis RCs, additional modifications are required in order to establish conditions that support bacterial replication, and transition to DCs.Vijay K. EedunuriYuntao ZhangChuanmin ChengLi ChenHuitao LiuAnders OmslandDan BoyleRoman R. GantaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Vijay K. Eedunuri Yuntao Zhang Chuanmin Cheng Li Chen Huitao Liu Anders Omsland Dan Boyle Roman R. Ganta Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
description	Abstract Ehrlichia chaffeensis, a tick-transmitted rickettsial bacterium, is the causative agent of human monocytic ehrlichiosis. Biochemical characterization of this and other related Rickettsiales remains a major challenge, as they require a host cell for their replication. We investigated the use of an axenic medium for E. chaffeensis growth, assessed by protein and DNA synthesis, in the absence of a host cell. E. chaffeensis organisms harvested from in vitro cultures grown in a vertebrate cell line were fractionated into infectious dense-core cells (DC) and the non-infectious replicating form, known as reticulate cells (RC) by renografin density gradient centrifugation and incubated in the axenic medium containing amino acids, nucleotides, and different energy sources. Bacterial protein and DNA synthesis were observed in RCs in response to glucose-6-phosphate, although adenosine triphosphate, alpha-ketoglutarate or sodium acetate supported protein synthesis. The biosynthetic activity could not be detected in DCs in the axenic medium. While the data demonstrate de novo protein and DNA synthesis under axenic conditions for E. chaffeensis RCs, additional modifications are required in order to establish conditions that support bacterial replication, and transition to DCs.
format	article
author	Vijay K. Eedunuri Yuntao Zhang Chuanmin Cheng Li Chen Huitao Liu Anders Omsland Dan Boyle Roman R. Ganta
author_facet	Vijay K. Eedunuri Yuntao Zhang Chuanmin Cheng Li Chen Huitao Liu Anders Omsland Dan Boyle Roman R. Ganta
author_sort	Vijay K. Eedunuri
title	Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
title_short	Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
title_full	Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
title_fullStr	Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
title_full_unstemmed	Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium
title_sort	protein and dna synthesis demonstrated in cell-free ehrlichia chaffeensis organisms in axenic medium
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/5d6953aeb10646b4ba0e1e46b4cb5c76
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Protein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic medium

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