Surface Proteome Analysis and Characterization of Surface Cell Antigen (Sca) or Autotransporter Family of Rickettsia typhi
| dc.contributor.author | Sears, Khandra T. | en |
| dc.contributor.author | Ceraul, Shane M. | en |
| dc.contributor.author | Gillespie, Joseph J. | en |
| dc.contributor.author | Allen, Edwin D., Jr. | en |
| dc.contributor.author | Popov, Vsevolod L. | en |
| dc.contributor.author | Ammerman, Nicole C. | en |
| dc.contributor.author | Rahman, M. Sayeedur | en |
| dc.contributor.author | Azad, Abdu F. | en |
| dc.date.accessioned | 2014-07-01T19:26:29Z | en |
| dc.date.available | 2014-07-01T19:26:29Z | en |
| dc.date.issued | 2012-08-09 | en |
| dc.description.abstract | Surface proteins of the obligate intracellular bacterium Rickettsia typhi, the agent of murine or endemic typhus fever, comprise an important interface for host-pathogen interactions including adherence, invasion and survival in the host cytoplasm. In this report, we present analyses of the surface exposed proteins of R. typhi based on a suite of predictive algorithms complemented by experimental surface-labeling with thiol-cleavable sulfo-NHS-SS-biotin and identification of labeled peptides by LC MS/MS. Further, we focus on proteins belonging to the surface cell antigen (Sca) autotransporter (AT) family which are known to be involved in rickettsial infection of mammalian cells. Each species of Rickettsia has a different complement of sca genes in various states; R. typhi, has genes sca1 thru sca5. In silico analyses indicate divergence of the Sca paralogs across the four Rickettsia groups and concur with previous evidence of positive selection. Transcripts for each sca were detected during infection of L929 cells and four of the five Sca proteins were detected in the surface proteome analysis. We observed that each R. typhi Sca protein is expressed during in vitro infections and selected Sca proteins were expressed during in vivo infections. Using biotin-affinity pull down assays, negative staining electron microscopy, and flow cytometry, we demonstrate that the Sca proteins in R. typhi are localized to the surface of the bacteria. All Scas were detected during infection of L929 cells by immunogold electron microscopy. Immunofluorescence assays demonstrate that Scas 1–3 and 5 are expressed in the spleens of infected Sprague-Dawley rats and Scas 3, 4 and 5 are expressed in cat fleas (Ctenocephalides felis). Sca proteins may be crucial in the recognition and invasion of different host cell types. In short, continuous expression of all Scas may ensure that rickettsiae are primed i) to infect mammalian cells should the flea bite a host, ii) to remain infectious when extracellular and iii) to infect the flea midgut when ingested with a blood meal. Each Sca protein may be important for survival of R. typhi and the lack of host restricted expression may indicate a strategy of preparedness for infection of a new host. | en |
| dc.description.sponsorship | The research presented in this article was supported by funds from the National Institutes of Health/National Institute of Allergy and Infectious Diseases (AI017828 to AFA). JJG is grateful for support from the National Institute of Allergy and Infectious Diseases (NIAID Contract No. HHSN272200900040C awarded to Bruno Sobral, Virginia Bioinformatics Institute at Virginia Tech). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.citation | Sears KT, Ceraul SM, Gillespie JJ, Allen ED Jr, Popov VL, et al. (2012) Surface Proteome Analysis and Characterization of Surface Cell Antigen (Sca) or Autotransporter Family of Rickettsia typhi. PLoS Pathog 8(8): e1002856. doi:10.1371/journal.ppat.1002856 | en |
| dc.identifier.doi | https://doi.org/10.1371/journal.ppat.1002856 | en |
| dc.identifier.issn | 1553-7374 | en |
| dc.identifier.uri | http://hdl.handle.net/10919/49257 | en |
| dc.identifier.url | http://www.plospathogens.org/article/info%3Adoi%2F10.1371%2Fjournal.ppat.1002856 | en |
| dc.language.iso | en_US | en |
| dc.publisher | Public Library of Science | en |
| dc.rights | Creative Commons Attribution 4.0 International | en |
| dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en |
| dc.subject | Fleas | en |
| dc.subject | L929 cells | en |
| dc.subject | Outer membrane proteins | en |
| dc.subject | Rickettsia rickettsii | en |
| dc.subject | Rickettsia | en |
| dc.title | Surface Proteome Analysis and Characterization of Surface Cell Antigen (Sca) or Autotransporter Family of Rickettsia typhi | en |
| dc.title.serial | PLoS Pathogens | en |
| dc.type | Article - Refereed | en |
| dc.type.dcmitype | Text | en |