Rapid, directed transport of DC-SIGN clusters in the plasma membrane

Ping, Liu; Weinreb, Violetta; Ridilla, Marc; Betts, Laurie; Patel, Pratik; Silva, Aravinda M. de; Thompson, Nancy L.; Jacobson, Ken

doi:10.1126/sciadv.aao1616

Cited by 12 publications

(20 citation statements)

References 61 publications

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“…In contrast to our model, Liu et al showed constitutive role of microtubule based retrograde transport of DC-SIGN nanoclusters to bring pathogens to the perinuclear region for antigen processing[21]. In this study, DC-SIGN nanoclusters were unladen with pathogen or attached to viral particles.…”

Section: Discussioncontrasting

confidence: 69%

“…In resting cell membranes, the DC-SIGN tetramer self assembles into nanodomains, which have been described to undergo Brownian diffusion as well as apparent directed transport in some contexts [17][18][19]. For instance, DC-SIGN nanodomain tracking studies have reported that this receptor can undergo rapid, linear transport in the plane of the plasma membrane, suggesting that some directed mobility of DC-SIGN is possible [12,18,20,21]. As previously mentioned, directed transport at immunological contact sites (e.g., T cell or B cell synapses) often involves cytoskeleton/motor protein dependent centripetal motion of receptors toward the center of the contact site [10].…”

Section: Evidence For Active Transport Of Dc-sign Into the C Albicanmentioning

confidence: 99%

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Dectin-1 mediated DC-SIGN Recruitment toCandida albicansContact Sites

Choraghe

Neumann²

2020

Preprint

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At host-pathogen contact sites with Candida albicans, Dectin-1 activates pro-inflammatory signaling, while DC-SIGN promotes adhesion to the fungal surface. We observed that Dectin-1 and DC-SIGN collaborate to enhance capture/retention of C. albicans under fluid shear culture conditions. Therefore, we devised a cellular model system wherein we could investigate the interaction between these two receptors during the earliest stages of host-pathogen interaction. In cells expressing both receptors, DC-SIGN was quickly recruited to contact sites (103.15% increase) but Dectin-1 did not similarly accumulate. Once inside the contact site, FRAP studies revealed a strong reduction in lateral mobility of DC-SIGN (but not Dectin-1), consistent with DC-SIGN engaging in multivalent adhesive binding interactions with cell wall mannoprotein ligands. Interestingly, in the absence of Dectin-1 co-expression, DC-SIGN recruitment to the contact was much poorer, only 35.04%. These data suggested that Dectin-1 promotes the active recruitment of DC-SIGN to the contact site. We proposed that Dectin-1 signaling activates the RHOA pathway, leading to actomyosin contractility that promotes DC-SIGN recruitment, perhaps via the formation of a centripetal ActoMyosin Flow (AMF) directed into the contact site. Indeed, RHOA pathway inhibitors significantly reduced Dectin-1 associated DC-SIGN recruitment to the contact site. We used agent based modeling to predict DC-SIGN transport kinetics with (Directed+Brownian) and without (Brownian) the hypothesized actomyosin flow-mediated transport. The Directed+Brownian transport model predicted a DC-SIGN contact site recruitment (108.72%), similar to that we observed experimentally under receptor co-expression. Brownian diffusive transport alone predicted contact site DC-SIGN recruitment of only 54.02%. However, this value was similar to experimentally observed recruitment in cells without Dectin-1 or treated with RHOA inhibitor, suggesting that it accurately predicted DC-SIGN recruitment when a contact site AMF would not be generated. TIRF microscopy of nascent cell contacts on glucan-coated glass revealed Dectin-1 dependent DC-SIGN and F-actin (LifeAct) recruitment kinetics to early-stage contact site membranes. DC-SIGN entry followed F-actin with a temporal lag of 8.35 +/- 4.57 seconds, but this correlation was disrupted by treatment with RHOA inhibitor. Thus, computational and experimental evidence provides support for the existence of a Dectin-1/RHOA-dependent AMF that produces a force to drive DC-SIGN recruitment to pathogen contact sites, resulting in improved pathogen capture and retention by immunocytes. These data suggest that the rapid collaborative response of Dectin-1 and DC-SIGN in early contact sties might be important for the efficient acquisition of yeast under flow conditions, such as those that prevail in circulation or mucocutaneous sites of infection.

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Section: Discussioncontrasting

confidence: 69%

Section: Evidence For Active Transport Of Dc-sign Into the C Albicanmentioning

confidence: 99%

Dectin-1 mediated DC-SIGN Recruitment toCandida albicansContact Sites

Choraghe

Neumann²

2020

Preprint

View full text Add to dashboard Cite

show abstract

“…In resting cell membranes, the DC-SIGN tetramer self assembles into nanodomains, which have been described to undergo Brownian diffusion as well as apparent directed transport in some contexts [ 17 , 18 , 19 ]. For instance, DC-SIGN nanodomain tracking studies have reported that this receptor can undergo rapid, linear transport in the plane of the plasma membrane, suggesting that some directed mobility of DC-SIGN is possible [ 12 , 18 , 20 , 21 ]. As previously mentioned, directed transport at immunological contact sites (e.g., T cell or B cell synapses) often involves cytoskeleton/motor protein-dependent centripetal motion of receptors toward the center of the contact site [ 10 ].…”

Section: Resultsmentioning

confidence: 99%

Dectin-1-Mediated DC-SIGN Recruitment to Candida albicans Contact Sites

Choraghe

Neumann

2021

Life

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At host–pathogen contact sites with Candida albicans, Dectin-1 activates pro-inflammatory signaling, while DC-SIGN promotes adhesion to the fungal surface. We observed that Dectin-1 and DC-SIGN collaborate to enhance capture/retention of C. albicans under fluid shear culture conditions. Therefore, we devised a cellular model system wherein we could investigate the interaction between these two receptors during the earliest stages of host–pathogen interaction. In cells expressing both receptors, DC-SIGN was quickly recruited to contact sites (103.15% increase) but Dectin-1 did not similarly accumulate. Once inside the contact site, FRAP studies revealed a strong reduction in lateral mobility of DC-SIGN (but not Dectin-1), consistent with DC-SIGN engaging in multivalent adhesive binding interactions with cell wall mannoprotein ligands. Interestingly, in the absence of Dectin-1 co-expression, DC-SIGN recruitment to the contact was much poorer—only 35.04%. These data suggested that Dectin-1 promotes the active recruitment of DC-SIGN to the contact site. We proposed that Dectin-1 signaling activates the RHOA pathway, leading to actomyosin contractility that promotes DC-SIGN recruitment, perhaps via the formation of a centripetal actomyosin flow (AMF) directed into the contact site. Indeed, RHOA pathway inhibitors significantly reduced Dectin-1-associated DC-SIGN recruitment to the contact site. We used agent-based modeling to predict DC-SIGN transport kinetics with (“Directed + Brownian”) and without (“Brownian”) the hypothesized actomyosin flow-mediated transport. The Directed + Brownian transport model predicted a DC-SIGN contact site recruitment (106.64%), similar to that we observed experimentally under receptor co-expression. Brownian diffusive transport alone predicted contact site DC-SIGN recruitment of only 55.60%. However, this value was similar to experimentally observed DC-SIGN recruitment in cells without Dectin-1 or expressing Dectin-1 but treated with RHOA inhibitor, suggesting that it accurately predicted DC-SIGN recruitment when a contact site AMF would not be generated. TIRF microscopy of nascent cell contacts on glucan-coated glass revealed Dectin-1-dependent DC-SIGN and F-actin (LifeAct) recruitment kinetics to early stage contact site membranes. DC-SIGN entry followed F-actin with a temporal lag of 8.35 ± 4.57 s, but this correlation was disrupted by treatment with RHOA inhibitor. Thus, computational and experimental evidence provides support for the existence of a Dectin-1/RHOA-dependent AMF that produces a force to drive DC-SIGN recruitment to pathogen contact sites, resulting in improved pathogen capture and retention by immunocytes. These data suggest that the rapid collaborative response of Dectin-1 and DC-SIGN in early contact sties might be important for the efficient acquisition of yeast under flow conditions, such as those that prevail in circulation or mucocutaneous sites of infection.

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“…The trajectories are then analysed, traditionally via mean squared displacement versus time plots (see below), to reveal the characteristics of the underlying motion. Using the SPT approach, it has been found that the diffusion behaviour of molecules in the cell membrane varies or regularly changes between multiple canonical patterns, namely free diffusion [30], confined diffusion [30], hop diffusion [31], channelled diffusion [32], stimulation-induced temporary arrest of lateral diffusion (STALL diffusion) [33] and directed motion [34]. The most common diffusion types seen are illustrated in Figure 2B.…”

Section: Single Particle Trackingmentioning

confidence: 99%

Revealing Plasma Membrane Nano-Domains with Diffusion Analysis Methods

et al. 2020

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Nano-domains are sub-light-diffraction-sized heterogeneous areas in the plasma membrane of cells, which are involved in cell signalling and membrane trafficking. Throughout the last thirty years, these nano-domains have been researched extensively and have been the subject of multiple theories and models: the lipid raft theory, the fence model, and the protein oligomerization theory. Strong evidence exists for all of these, and consequently they were combined into a hierarchal model. Measurements of protein and lipid diffusion coefficients and patterns have been instrumental in plasma membrane research and by extension in nano-domain research. This has led to the development of multiple methodologies that can measure diffusion and confinement parameters including single particle tracking, fluorescence correlation spectroscopy, image correlation spectroscopy and fluorescence recovery after photobleaching. Here we review the performance and strengths of these methods in the context of their use in identification and characterization of plasma membrane nano-domains.

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Rapid, directed transport of DC-SIGN clusters in the plasma membrane

Abstract: Very rapid, directed movement of pathogen receptors in the plasma membrane is associated with MT close to the inner leaflet.

Cited by 12 publications

References 61 publications

Dectin-1 mediated DC-SIGN Recruitment toCandida albicansContact Sites

Dectin-1 mediated DC-SIGN Recruitment toCandida albicansContact Sites

Dectin-1-Mediated DC-SIGN Recruitment to Candida albicans Contact Sites

Revealing Plasma Membrane Nano-Domains with Diffusion Analysis Methods

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