Ehrlichia chaffeensis Type 1 Secreted Effector TRP47 is a Novel SUMOylated Nucleomodulin
| dc.contributor.advisor | McBride, Jere W | |
| dc.contributor.committeeMember | Walker, David H | |
| dc.contributor.committeeMember | Sahni, Sanjeev K | |
| dc.creator | Milligan, Sarah L | |
| dc.creator.orcid | 0000-0002-6116-1372 | |
| dc.date.accessioned | 2019-03-13T20:47:26Z | |
| dc.date.available | 2019-03-13T20:47:26Z | |
| dc.date.created | 2015-12 | |
| dc.date.submitted | December 2015 | |
| dc.date.updated | 2019-03-13T20:47:26Z | |
| dc.description.abstract | Ehrlichia chaffeensis secretes immunodominant and immunoprotective protein effectors into the host cell using a type I secretion system. The best-characterized of these effectors are the tandem repeat proteins (TRPs). In this study, E. chaffeensis TRP47 was identified as a target for host cell SUMOylation and as a novel nucleomodulin with DNA-binding activity and host nuclear localization. Using a microfluidic chip peptide array, a SUMOylation site in the TRP47 N-terminal domain was identified. A SUMO-2 modification was further confirmed using an in vitro SUMOylation assay with recombinant TRP47, and native polySUMOylated TRP47 was immunoprecipitated from E. chaffeensis-infected cell lysates. TRP47 was detected in host cell nuclei primarily at 24 and 48 hours post-infection by immunofluorescence microscopy. A domain in the N-terminal portion of the protein containing a variant MYND-binding motif was identified as the region responsible for TRP47 nuclear localization using ectopically expressed GFP-tagged TRP47 truncation constructs. Recombinant TRP47 also binds human genomic DNA in an electrophoretic mobility shift assay. Taken together, these data demonstrate that E. chaffeensis TRP47 is a nucleomodulin with DNA-binding activity and a non-canonical nuclear localization signal, and is post-translationally modified by SUMOylation, which is a novel modification among bacteria. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | https://hdl.handle.net/2152.3/11208 | |
| dc.subject | Ehrlichia | |
| dc.subject | nucleomodulin | |
| dc.subject | SUMOylation | |
| dc.title | Ehrlichia chaffeensis Type 1 Secreted Effector TRP47 is a Novel SUMOylated Nucleomodulin | |
| dc.type | Thesis | |
| dc.type.material | text | |
| thesis.degree.department | Experimental Pathology | |
| thesis.degree.grantor | The University of Texas Medical Branch at Galveston | |
| thesis.degree.level | Masters | |
| thesis.degree.name | Experimental Pathology (Masters) |