<i>Anaplasma phagocytophilum</i>Rab10-dependent parasitism of the<i>trans</i>-Golgi network is critical for completion of the infection cycle

Truchan, Hilary K.; VieBrock, Lauren; Cockburn, Chelsea L.; Ojogun, Nore; Griffin, Brian P.; Wijesinghe, Dayanjan S.; Chalfant, Charles E.; Carlyon, Jason A.

doi:10.1111/cmi.12500

Cited by 31 publications

(72 citation statements)

References 65 publications

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“…), and primate RF/6A endothelial cells (both species) (Goodman et al, 1996 ; Woldehiwet et al, 2002 ; Munderloh et al, 2004 ; Zivkovic et al, 2009 ). RF/6A cells are particularly useful models for studying the cellular microbiology of these bacteria because they are large and flat, making them ideal for imaging (Munderloh et al, 2004 ; Sukumaran et al, 2011 ; Beyer et al, 2014 ; Truchan et al, 2016 ). Moreover, A. phagocytophilum and A. marginale have been detected in endothelial cells of tissue sections recovered from experimentally infected animals (Herron et al, 2005 ; Wamsley et al, 2011 ).…”

Section: Introductionmentioning

confidence: 99%

The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum

Truchan

Cockburn

Hebert

et al. 2016

Front. Cell. Infect. Microbiol.

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The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale invades bovine erythrocytes. Evidence suggests that both species may also infect endothelial cells in vivo. In mammalian and arthropod host cells, A. phagocytophilum and A. marginale reside in host cell derived pathogen-occupied vacuoles (POVs). While it was recently demonstrated that the A. phagocytophilum-occupied vacuole (ApV) intercepts membrane traffic from the trans-Golgi network, it is unclear if it or the A. marginale-occupied vacuole (AmV) interacts with other secretory organelles. Here, we demonstrate that the ApV and AmV extensively interact with the host endoplasmic reticulum (ER) in endothelial, myeloid, and/or tick cells. ER lumen markers, calreticulin, and protein disulfide isomerase, and the ER membrane marker, derlin-1, were pronouncedly recruited to the peripheries of both POVs. ApV association with the ER initiated early and continued throughout the infection cycle. Both the ApV and AmV interacted with the rough ER and smooth ER. However, only derlin-1-positive rough ER derived vesicles were delivered into the ApV lumen where they localized with intravacuolar bacteria. Transmission electron microscopy identified multiple ER-POV membrane contact sites on the cytosolic faces of both species' vacuoles that corresponded to areas on the vacuoles' lumenal faces where intravacuolar Anaplasma organisms closely associated. A. phagocytophilum is known to hijack Rab10, a GTPase that regulates ER dynamics and morphology. Yet, ApV-ER interactions were unhindered in cells in which Rab10 had been knocked down, demonstrating that the GTPase is dispensable for the bacterium to parasitize the ER. These data establish the ApV and AmV as pathogen-host interfaces that directly engage the ER in vertebrate and invertebrate host cells and evidence the conservation of ER parasitism between two Anaplasma species.

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Section: Introductionmentioning

confidence: 99%

The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum

Truchan

Cockburn

Hebert

et al. 2016

Front. Cell. Infect. Microbiol.

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“…Similar to A. phagocytophilum, the AmVs engaged the ER and the Golgi apparatus early in infection and maintained association throughout the course of infection (29,38,39). The ER and the Golgi apparatus are nutrient-rich compartments of the secretory pathway and likely serve as a rich source of glycoprotein and lipids, which can be degraded to supply amino acids, fatty acids, nitrogen, carbon, and energy to auxotrophic A. marginale (40,41).…”

Section: Discussionmentioning

confidence: 99%

“…The dynamics of transport in the secretory pathway are complex and involve many molecules that are targeted by organisms that parasitize these nutrient-rich compartments. These molecules include Rab10 targeted by A. phagocytophilum for hijacking secretory vesicles from the trans-Golgi network (38) and Rab1 used by Legionella pneumophila to engage traffic from the ER (16).…”

Section: Discussionmentioning

confidence: 99%

Anaplasma marginale Actively Modulates Vacuolar Maturation during Intracellular Infection of Its Tick Vector, Dermacentor andersoni

Magunda

Thompson

Schneider

et al. 2016

Appl Environ Microbiol

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Tick-borne transmission of bacterial pathogens in the order Rickettsiales is responsible for diverse infectious diseases, many of them severe, in humans and animals. Transmission dynamics differ among these pathogens and are reflected in the pathogenvector interaction. Anaplasma marginale has been shown to establish and maintain infectivity within Dermacentor spp. for weeks to months while escaping the complex network of vacuolar peptidases that are responsible for digestion of the tick blood meal. How this prolonged maintenance of infectivity in a potentially hostile environment is achieved has been unknown. Using the natural vector Dermacentor andersoni, we demonstrated that A. marginale-infected tick vacuoles (AmVs) concurrently recruit markers of the early endosome (Rab5), recycling endosome (Rab4 and Rab11), and late endosome (Rab7), are maintained near neutral pH, do not fuse with lysosomes, exclude the protease cathepsin L, and engage the endoplasmic reticulum and Golgi apparatus for up to 21 days postinfection. Maintenance of this safe vacuolar niche requires active A. marginale protein synthesis; in its absence, the AmVs mature into acidic, protease-active phagolysosomes. Identification of this bacterially directed modeling of the tick midgut endosome provides a mechanistic basis for examination of the differences in transmission efficiency observed among A. marginale strains and among vector populations. T icks are biological vectors of the obligate intracellular bacteria of the order Rickettsiales, including pathogens that cause severe diseases in humans and animals (1). Transmission dynamics determine the epidemiologic features of a given disease and vary among the pathogen-vector pairs. For example, Rickettsia rickettsii, the causative agent of Rocky Mountain spotted fever, is maintained in the tick vector Dermacentor andersoni through transovarial transmission, and thus each new generation of larval ticks is infected. Additionally, infection is maintained with each successive molt, circumventing the need for feeding on an infected mammalian host to maintain the pathogen through time. In contrast, Anaplasma marginale, responsible for anaplasmosis in ruminants, must be acquired and transstadially transmitted by each new generation of adult D. andersoni ticks (2). It is not transovarially transmitted, and the larvae and nymphs of D. andersoni are not exposed to A. marginale because they feed preferentially on small mammals. Adult male ticks feed preferentially on cattle, move between hosts while seeking mates, and thus are the epidemiologically important transmission vector. Consequently, A. marginale must persist in the tick for the period of time necessary for an adult tick to ingest two separate blood meals, which depending on conditions, may occur in separate seasons. Experimentally infected adult male D. andersoni ticks are able to transmit A. marginale for up to 26 days after acquisition feeding (2-4). The mechanisms by which pathogens maintain long-term infections within ticks remain po...

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“…A confluent flask of untransfected HeLa cells was used as a control. At 4‐ or 16‐hr posttransfection, the cells were washed twice with cold phosphate buffered saline prior to processing for density gradient centrifugation and trichloroacetic acid precipitation as previously described (Truchan et al, ). Equal volumes of fractions 1–9 were immunoblotted using GM130 (1:500, BD Biosciences), calreticulin (1:4000, Sigma Aldrich), and GFP (1:1000, Thermo Fisher) antibodies.…”

Section: Methodsmentioning

confidence: 99%

“…At 4-or 16-hr posttransfection, the cells were washed twice with cold phosphate buffered saline prior to processing for density gradient centrifugation and trichloroacetic acid precipitation as previously described (Truchan et al, 2016). Equal volumes of fractions 1-9…”

Section: Density Gradient Centrifugationmentioning

confidence: 99%

Orientia tsutsugamushiAnk9 is a multifunctional effector that utilizes a novel GRIP-like Golgi localization domain for Golgi-to-endoplasmic reticulum trafficking and interacts with host COPB2

Beyer

Rodino

VieBrock

et al. 2017

Cellular Microbiology

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SUMMARY Orientia tsutsugamushi causes scrub typhus, a potentially fatal infection that afflicts one million people annually. This obligate intracellular bacterium boasts one of the largest microbial arsenals of ankyrin repeat-containing protein (Ank) effectors, most of which target the endoplasmic reticulum (ER) by undefined mechanisms. Ank9 is the only one proven to function during infection. Here, we demonstrate that Ank9 bears a motif that mimics the GRIP domain of eukaryotic golgins and is necessary and sufficient for its Golgi localization. Ank9 reaches the ER exclusively by retrograde trafficking from the Golgi. Consistent with this observation, it binds COPB2, a host protein that mediates Golgi-to-ER transport. Ank9 destabilizes the Golgi and ER in a Golgi-localization domain (GLD)-dependent manner and induces the ATF4-dependent unfolded protein response. The Golgi is also destabilized in cells infected with O. tsutsugamushi or treated with COPB2 siRNA. COPB2 reduction and/or the cellular events that it invokes, such as Golgi destabilization, benefit Orientia replication. Thus, Ank9/bacterial negative modulation of COPB2 might contribute to the bacterium’s intracellular replication. This report identifies a novel microbial GLD, links Ank9 to the ability of O. tsutsugamushi to perturb Golgi structure, and describes the first mechanism by which any Orientia effector targets the secretory pathway.

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Anaplasma phagocytophilumRab10-dependent parasitism of thetrans-Golgi network is critical for completion of the infection cycle

Cited by 31 publications

References 65 publications

The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum

The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum

Anaplasma marginale Actively Modulates Vacuolar Maturation during Intracellular Infection of Its Tick Vector, Dermacentor andersoni

Orientia tsutsugamushiAnk9 is a multifunctional effector that utilizes a novel GRIP-like Golgi localization domain for Golgi-to-endoplasmic reticulum trafficking and interacts with host COPB2

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