Interplay between Clathrin and Rab5 Controls the Early Phagocytic Trafficking and Intracellular Survival of Brucella abortus within HeLa cells

Lee, Jin Ju; Kim, Dae Geun; Kim, Dong Hyeok; Simborio, Hannah Leah Tadeja; Min, Wongi; Lee, Hu Jang; Her, Moon; Jung, Suk-Chan; Watarai, Masahisa; Kim, Suk

doi:10.1074/jbc.m113.491555

Cited by 29 publications

(31 citation statements)

References 49 publications

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“…The usurpation of CME during the internalization/invasion process is only recently being recognized to be employed by several bacteria, best studied in Listeria monocytogenes infection [129, 130]. However, only one study on Brucella abortus , a pathogen also residing in a vacuole during intracellular lifestyle, demonstrated the association of clathrin with the Brucella -containing vacuole [131]. We now show such an association with CLTA also for STM (Figure 7).…”

Section: Discussionsupporting

confidence: 67%

A trafficome-wide RNAi screen reveals deployment of early and late secretory host proteins and the entire late endo-/lysosomal vesicle fusion machinery by intracellularSalmonella

Kehl

Göser

Reuter

et al. 2019

Preprint

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38 The intracellular lifestyle of Salmonella enterica is characterized by the formation of a 39 replication-permissive membrane-bound niche, the Salmonella-containing vacuole (SCV). A 40 further consequence of the massive remodeling of the host cell endosomal system, intracellular 41 Salmonella establish a unique network of various Salmonella-induced tubules (SIT). The 42 bacterial repertoire of effector proteins required for the establishment for one type of these SIT, 43 the Salmonella-induced filaments (SIF), is rather well-defined. However, the corresponding 44 host cell proteins are still poorly understood. To identify host factors required for the formation 45 of SCV and SIF, we performed a sub-genomic RNAi screen. The analyses comprised high-46 resolution live cell imaging to score effects on SIF induction, dynamics and morphology. The 47 hits of our functional RNAi screen comprise: i) The late endo-/lysosomal SNARE (soluble N-48 ethylmaleimide-sensitive factor attachment protein receptor) complex, consisting of STX7, 49 STX8, VTI1B, and VAMP7 or VAMP8, this is, in conjunction with RAB7 and the homotypic 50 fusion and protein sorting (HOPS) tethering complex, a complete vesicle fusion machinery. ii) 51 Novel interactions with the early secretory GTPases RAB1A and RAB1B, possibly providing 52 a link to coat protein complex I (COPI) vesicles and reinforcing recently identified ties to the 53 endoplasmic reticulum. iii) New connections to the late secretory pathway and/or the recycling 54 endosome via the GTPases RAB3A, RAB8A, and RAB8B and the SNAREs VAMP2, VAMP3, 55 and VAMP4. iv) An unprecedented involvement of clathrin-coated structures. The resulting set 56 of hits allowed to characterize completely new host factor interactions, and strengthen 57 observations from several previous studies. 58 Author Summary59 The facultative intracellular pathogen Salmonella enterica serovar Typhimurium induces the 60 reorganization of the endosomal system of mammalian host cells. This activity is dependent on 61 translocated effector proteins of the pathogen. The host cells factors required for endosomal 18.11.2019 Host factors for SIF formation 4 62 remodeling are only partially known. To identify such factors for formation and dynamics of 63 endosomal compartments in Salmonella-infected cell, we performed a live cell imaging-based 64 RNAi screen a to investigate the role of 496 mammalian proteins involved in cellular logistics. 65 We identified that endosomal remodeling by intracellular Salmonella dependent on host factors 66 in following functional classes: i) the late endo-/lysosomal SNARE (soluble N-ethylmaleimide-67 sensitive factor attachment protein receptor) complex, ii) the early secretory pathway, 68 represented by regulators GTPases RAB1A and RAB1B, iii) the late secretory pathway and/or 69 recycling endosomes represented by GTPases RAB3A, RAB8A, RAB8B, and the SNAREs 70 VAMP2, VAMP3, and VAMP4, and iv) clathrin-coated structures. The identification of these 71 new host factors provides further evidence ...

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Section: Discussionsupporting

confidence: 67%

A trafficome-wide RNAi screen reveals deployment of early and late secretory host proteins and the entire late endo-/lysosomal vesicle fusion machinery by intracellularSalmonella

Kehl

Göser

Reuter

et al. 2019

Preprint

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“…This assay was performed as previously reported (Kim, Watanabe, et al, ; Lee et al, ). Briefly, macrophages (10 6 cells) were seeded into 96‐well plates, incubated for 24 hr at 37 °C in an incubator with 5% CO 2 and then infected with 10 7 CFU of virulent B. abortus .…”

Section: Methodsmentioning

confidence: 99%

“…The colocalisation of Brucella-containing phagosomes (BCPs) with LAMP-1 was evaluated as described in previous reports (Kim, Kurokawa, et al, 2004;Lee et al, 2013). Briefly, RAW 264.7 cells were treated with the Lcn2 siRNA prior to infection.…”

Section: Lamp-1 Colocalisationmentioning

confidence: 99%

Lipocalin 2 (Lcn2) interferes with iron uptake byBrucella abortusand dampens immunoregulation during infection of RAW 264.7 macrophages

Hop

Arayan

Huy

et al. 2017

Cellular Microbiology

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Lipocalin 2 (Lcn2) is an important innate immunity component against bacterial pathogens. In this study, we report that Lcn2 is induced by Brucella (B.) abortus infection and significantly contributes to the restriction of intracellular survival of Brucella in macrophages. We found that Lcn2 prevented iron uptake by B. abortus through two distinct mechanisms. First, Lcn2 is secreted to capture bacterial siderophore(s) and abrogate iron import by Brucella. Second, Lcn2 decreases the intracellular iron levels during Brucella infection, which probably deprives the invading Brucella of the iron source needed for growth. Suppression of Lcn2 signalling resulted in a marked induction of anti-inflammatory cytokine, interleukin 10, which was shown to play a major role in Lcn2-induced antibrucella immunity. Similarly, interleukin 6 was also found to be increased when Lcn2 signalling is abrogated; however, this induction was thought to be an alternative pathway that rescues the cell from infection when the effective Lnc2 pathway is repressed. Furthermore, Lcn2 deficiency also caused a marked decrease in brucellacidal effectors, such as reactive oxygen species and nitric oxide but not the phagolysosome fusion. Taken together, our results indicate that Lcn2 is required for the efficient restriction of intracellular B. abortus growth that is through limiting iron acquisition and shifting cells to pro-inflammatory brucellacidal activity in murine macrophages.

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“…are facultative intracellular Gramnegative bacteria that cause brucellosis in a variety of mammalian hosts, particularly, they cause more than 500,000 new human cases reported annually (1). They can prevent phagosome maturation by not fully understood mechanisms, leading to successful survival within professional and nonprofessional phagocytes (2,3). Vaccination seems to be a predominant manner for the control of infectious diseases; however, there is no 100% efficacious vaccine for brucellosis so far.…”

Section: Introductionmentioning

confidence: 99%

Interleukin 10 suppresses lysosome-mediated killing of Brucella abortus in cultured macrophages

Hop¹,

Reyes²,

Huy³

et al. 2018

Journal of Biological Chemistry

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Interplay between Clathrin and Rab5 Controls the Early Phagocytic Trafficking and Intracellular Survival of Brucella abortus within HeLa cells

Cited by 29 publications

References 49 publications

A trafficome-wide RNAi screen reveals deployment of early and late secretory host proteins and the entire late endo-/lysosomal vesicle fusion machinery by intracellularSalmonella

A trafficome-wide RNAi screen reveals deployment of early and late secretory host proteins and the entire late endo-/lysosomal vesicle fusion machinery by intracellularSalmonella

Lipocalin 2 (Lcn2) interferes with iron uptake byBrucella abortusand dampens immunoregulation during infection of RAW 264.7 macrophages

Interleukin 10 suppresses lysosome-mediated killing of Brucella abortus in cultured macrophages

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