In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice.
Analysis of the transcriptome of Borrelia burgdorferi, the causative agent of Lyme disease, during infection has proven difficult due to the low spirochete loads in the mammalian tissues. To overcome this challenge, we have developed an In Vivo Expression Technology (IVET) system for identification...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC3757035?pdf=render |
| _version_ | 1818028727317037056 |
|---|---|
| author | Tisha Choudhury Ellis Sunny Jain Angelika K Linowski Kelli Rike Aaron Bestor Patricia A Rosa Micah Halpern Stephanie Kurhanewicz Mollie W Jewett |
| author_facet | Tisha Choudhury Ellis Sunny Jain Angelika K Linowski Kelli Rike Aaron Bestor Patricia A Rosa Micah Halpern Stephanie Kurhanewicz Mollie W Jewett |
| author_sort | Tisha Choudhury Ellis |
| collection | DOAJ |
| description | Analysis of the transcriptome of Borrelia burgdorferi, the causative agent of Lyme disease, during infection has proven difficult due to the low spirochete loads in the mammalian tissues. To overcome this challenge, we have developed an In Vivo Expression Technology (IVET) system for identification of B. burgdorferi genes expressed during an active murine infection. Spirochetes lacking linear plasmid (lp) 25 are non-infectious yet highly transformable. Mouse infection can be restored to these spirochetes by expression of the essential lp25-encoded pncA gene alone. Therefore, this IVET-based approach selects for in vivo-expressed promoters that drive expression of pncA resulting in the recovery of infectious spirochetes lacking lp25 following a three week infection in mice. Screening of approximately 15,000 clones in mice identified 289 unique in vivo-expressed DNA fragments from across all 22 replicons of the B. burgdorferi B31 genome. The in vivo-expressed candidate genes putatively encode proteins in various functional categories including antigenicity, metabolism, motility, nutrient transport and unknown functions. Candidate gene bbk46 on essential virulence plasmid lp36 was found to be highly induced in vivo and to be RpoS-independent. Immunocompetent mice inoculated with spirochetes lacking bbk46 seroconverted but no spirochetes were recovered from mouse tissues three weeks post inoculation. However, the bbk46 gene was not required for B. burgdorferi infection of immunodeficient mice. Therefore, through an initial IVET screen in B. burgdorferi we have identified a novel in vivo-induced virulence factor critical for the ability of the spirochete to evade the humoral immune response and persistently infect mice. |
| first_indexed | 2024-12-10T05:08:23Z |
| format | Article |
| id | doaj.art-7087bcf6f2fa4219bbe2622d53bf7be2 |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-10T05:08:23Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-7087bcf6f2fa4219bbe2622d53bf7be22022-12-22T02:01:09ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742013-01-0198e100356710.1371/journal.ppat.1003567In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice.Tisha Choudhury EllisSunny JainAngelika K LinowskiKelli RikeAaron BestorPatricia A RosaMicah HalpernStephanie KurhanewiczMollie W JewettAnalysis of the transcriptome of Borrelia burgdorferi, the causative agent of Lyme disease, during infection has proven difficult due to the low spirochete loads in the mammalian tissues. To overcome this challenge, we have developed an In Vivo Expression Technology (IVET) system for identification of B. burgdorferi genes expressed during an active murine infection. Spirochetes lacking linear plasmid (lp) 25 are non-infectious yet highly transformable. Mouse infection can be restored to these spirochetes by expression of the essential lp25-encoded pncA gene alone. Therefore, this IVET-based approach selects for in vivo-expressed promoters that drive expression of pncA resulting in the recovery of infectious spirochetes lacking lp25 following a three week infection in mice. Screening of approximately 15,000 clones in mice identified 289 unique in vivo-expressed DNA fragments from across all 22 replicons of the B. burgdorferi B31 genome. The in vivo-expressed candidate genes putatively encode proteins in various functional categories including antigenicity, metabolism, motility, nutrient transport and unknown functions. Candidate gene bbk46 on essential virulence plasmid lp36 was found to be highly induced in vivo and to be RpoS-independent. Immunocompetent mice inoculated with spirochetes lacking bbk46 seroconverted but no spirochetes were recovered from mouse tissues three weeks post inoculation. However, the bbk46 gene was not required for B. burgdorferi infection of immunodeficient mice. Therefore, through an initial IVET screen in B. burgdorferi we have identified a novel in vivo-induced virulence factor critical for the ability of the spirochete to evade the humoral immune response and persistently infect mice.http://europepmc.org/articles/PMC3757035?pdf=render |
| spellingShingle | Tisha Choudhury Ellis Sunny Jain Angelika K Linowski Kelli Rike Aaron Bestor Patricia A Rosa Micah Halpern Stephanie Kurhanewicz Mollie W Jewett In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. PLoS Pathogens |
| title | In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. |
| title_full | In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. |
| title_fullStr | In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. |
| title_full_unstemmed | In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. |
| title_short | In vivo expression technology identifies a novel virulence factor critical for Borrelia burgdorferi persistence in mice. |
| title_sort | in vivo expression technology identifies a novel virulence factor critical for borrelia burgdorferi persistence in mice |
| url | http://europepmc.org/articles/PMC3757035?pdf=render |
| work_keys_str_mv | AT tishachoudhuryellis invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT sunnyjain invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT angelikaklinowski invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT kellirike invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT aaronbestor invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT patriciaarosa invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT micahhalpern invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT stephaniekurhanewicz invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice AT molliewjewett invivoexpressiontechnologyidentifiesanovelvirulencefactorcriticalforborreliaburgdorferipersistenceinmice |