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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| Format: | Article |
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Nature Portfolio
2018-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-018-27574-z |
| _version_ | 1818995930592968704 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-20T21:21:40Z |
| format | Article |
| id | doaj.art-5d6953aeb10646b4ba0e1e46b4cb5c76 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T21:21:40Z |
| publishDate | 2018-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-5d6953aeb10646b4ba0e1e46b4cb5c762022-12-21T19:26:15ZengNature PortfolioScientific Reports2045-23222018-06-018111210.1038/s41598-018-27574-zProtein and DNA synthesis demonstrated in cell-free Ehrlichia chaffeensis organisms in axenic mediumVijay K. Eedunuri0Yuntao Zhang1Chuanmin Cheng2Li Chen3Huitao Liu4Anders Omsland5Dan Boyle6Roman R. Ganta7Center of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityCenter of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityCenter of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityCenter of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityCenter of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityPaul G. Allen School for Global Animal Health, Washington State UniversityDivision of Biology, Kansas State UniversityCenter of Excellence for Vector-Borne Diseases, Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State UniversityAbstract 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.https://doi.org/10.1038/s41598-018-27574-z |
| spellingShingle | 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 Scientific Reports |
| title | 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_short | 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 |
| url | https://doi.org/10.1038/s41598-018-27574-z |
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