Biology subjects

Tran, C. J.

Publications and source records attributed to Tran, C. J..

4 recordsLinked to original sources

bioRxiv2024

The Rickettsia actin-based motility effectors RickA and Sca2 contribute differently to cell-to-cell spread and pathogenicity

Rickettsia parkeri is an obligate intracellular, tick-borne bacterial pathogen that can cause eschar-associated rickettsiosis in humans. R. parkeri invades host cells, escapes from vacuoles into the cytosol, and undergoes two independent modes of actin-based motility mediated by effectors RickA or Sca2. Actin-based motility of R. parkeri enables bacteria to enter protrusions of the host cell plasma membrane that are engulfed by neighboring host cells. However, whether and how RickA and Sca2 independently contribute to cell-to-cell spread in vitro or pathogenicity in vivo has been unclear. Using live cell imaging of rickA::Tn and sca2::Tn mutants, we discovered both RickA and Sca2 contribute to different modes of cell-to-cell spread. Compared with Sca2-spread, RickA-spread involves the formation of longer protrusions that exhibit larger fluctuations in length and take a longer time to be engulfed into neighboring cells. We further compared the roles of RickA and Sca2 in vivo following intradermal infection of Ifnar1-/-; Ifngr1-/- double-knockout mice, which exhibit eschars and succumb to infection with wild-type R. parkeri. We observed that RickA is important for severe eschar formation, whereas Sca2 contributes to larger foci of infection in the skin and dissemination from the skin to the internal organs. Our results suggest that actin-based motility effectors RickA and Sca2 drive two distinct forms of cell-to-cell spread and contribute differently to pathogenicity in the mammalian host.

microbiology↗

bioRxiv2024

The Chlamydia effector IncE employs two short linear motifs to reprogram host vesicle trafficking

Chlamydia trachomatis, a leading cause of bacteria sexually transmitted infections, creates a specialized intracellular replicative niche by translocation and insertion of a diverse array of effectors (Incs) into the inclusion membrane. Here, we characterize IncE, a multi-functional Inc that encodes two non-overlapping short linear motifs (SLiMs) within its short cytosolic C-terminus. The proximal SLiM mimics an R-SNARE motif to recruit syntaxin (STX) 7 and 12-containing vesicles to the inclusion. The distal SLiM mimics the Sorting Nexin (SNX) 5 and 6 cargo binding site to recruit SNX6-containing vesicles to the inclusion. By simultaneously binding to two distinct vesicle classes, IncE reprograms host cell trafficking to promote the formation of a class of hybrid vesicles at the inclusion that are required for C. trachomatis intracellular development. Our work suggests that Incs may have evolved SLiMs to facilitate rapid evolution in a limited protein space to disrupt host cell processes.

microbiology↗

bioRxiv2021

A patatin-like phospholipase mediates Rickettsia parkeri escape from host membranes

Spotted fever group Rickettsia species are arthropod-borne obligate intracellular bacteria that can cause mild to severe human disease. These bacteria invade host cells, replicate in the cell cytosol, and then spread from cell to cell. To access the host cytosol and avoid detection by immune surveillance mechanisms, these pathogens must have evolved efficient ways to escape membrane-bound vacuoles. Although Rickettsia are predicted to express factors that disrupt host membranes, little is known about how and when these proteins function during infection. Here, we investigated the role of a Rickettsia patatin-like phospholipase A2 enzyme (Pat1) during host cell infection by characterizing a Rickettsia parkeri mutant with a transposon insertion in the pat1 gene. We show that Pat1 is important for infection in a mouse model and in host cells. We further show that Pat1 is critical for efficiently escaping from the single and double membrane-bound vacuoles into the host cytosol, and for avoiding host galectins that mark damaged membranes. In the host cytosol, Pat1 is important for avoiding host polyubiquitin, preventing recruitment of autophagy receptor p62, and promoting actin-based motility and cell-cell spread. Our results show that Pat1 plays critical roles in escaping host membranes and promoting cell-cell spread during R. parkeri infection and suggest diverse roles for patatin-like phospholipases in facilitating microbial infection. ImportanceSpotted fever group Rickettsia are bacteria that reside in ticks and can be transmitted to mammalian hosts, including humans. Severe disease is characterized by high fever, headache, and rash, and results in occasional mortality despite available treatment. Rickettsia interact with host cell membranes while invading cells, escaping into the cytosol, and evading cellular defenses. Bacterial phospholipase enzymes have been proposed as critical factors for targeting host cell membranes, however the specific roles of rickettsial phospholipases are not well defined. We investigated the contribution of one conserved patatin-like phospholipase, Pat1, in Rickettsia parkeri. We observed that Pat1 is important for virulence in an animal model. Moreover, Pat1 plays critical roles in host cells by facilitating access to the cell cytosol, inhibiting detection by host defense pathways, and promoting cell-cell spread. Our study indicates that Pat1 performs several critical functions, suggesting a broad role for phospholipases throughout the Rickettsia lifecycle.

microbiology↗

bioRxiv2020

Rickettsia parkeri Sca2 promotes dissemination in an intradermal infection mouse model

Rickettsia are arthropod-borne pathogens that cause severe human disease worldwide. The spotted fever group (SFG) pathogen Rickettsia parkeri elicits skin lesion (eschar) formation in humans after tick bite. However, intradermal inoculation of inbred mice with millions of bacteria fails to elicit eschar formation or disseminated disease, hindering investigations into understanding eschar-associated rickettsiosis. Here, we report that intradermal infection of mice deficient for both interferon receptors (Ifnar-/-Ifngr-/-) with R. parkeri causes eschar formation, recapitulating the hallmark clinical feature of human disease. Intradermal infection with doses that recapitulate tick infestation caused eschar formation and lethality, including with as few as 10 bacteria. Using this model, we found that the actin-based motility protein Sca2 is required for R. parkeri dissemination from the skin to internal organs and for causing lethal disease, and that the abundant R. parkeri outer membrane protein OmpB contributes to eschar formation. We also found that immunizing mice with sca2 and ompB mutant R. parkeri protects against subsequent rechallenge with wild-type bacteria, revealing live-attenuated vaccine candidates. Thus, interferon receptor-deficient mice are a tractable model to investigate rickettsiosis, bacterial virulence factors, and immunity. Our results suggest that differences in interferon signaling in the skin between mice and humans may explain the discrepancy in susceptibility to SFG Rickettsia.

microbiology↗