Secretory Vesicle Clustering in Fungal Filamentous Cells Does Not Require Directional Growth

Silva, Patrícia M.; Puerner, Charles; Seminara, Agnese; Bassilana, Martine; Arkowitz, Robert A.

doi:10.1016/j.celrep.2019.07.062

Cited by 17 publications

(29 citation statements)

References 105 publications

(141 reference statements)

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“…When a new polarity site needs to be established in an already asymmetric cell, experimentally increasing the level of active Cdc42p on the plasma membrane results in mislocalization of the exocyst subunit Sec3p and an altered clustering of secretory vesicles. This new cluster of secretory vesicles is highly dynamic, until a new growth site can be established [342]. Cdc42p also promotes polarized recruitment of several polarisome components, including Spa2p and the formin Bni1p in C. albicans.…”

Section: Polarized Secretion In Candida Albicans Hyphaementioning

confidence: 99%

The Role of Secretory Pathways in Candida albicans Pathogenesis

Rollenhagen

Mamtani

et al. 2020

JoF

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Candida albicans is a fungus that is a commensal organism and a member of the normal human microbiota. It has the ability to transition into an opportunistic invasive pathogen. Attributes that support pathogenesis include secretion of virulence-associated proteins, hyphal formation, and biofilm formation. These processes are supported by secretion, as defined in the broad context of membrane trafficking. In this review, we examine the role of secretory pathways in Candida virulence, with a focus on the model opportunistic fungal pathogen, Candida albicans.

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Section: Polarized Secretion In Candida Albicans Hyphaementioning

confidence: 99%

The Role of Secretory Pathways in Candida albicans Pathogenesis

Rollenhagen

Mamtani

et al. 2020

JoF

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“…Figure 5 d shows the probe prior to PDMS rupture and subsequent to exiting from the PDMS. Experiments were carried out with a range of probe displacement rates encompassing that of the filament extension rates (~ 0.3 μm/min [ 33 ]), when scaled down to the filament diameter. Figure 6 a shows an example of such a force versus displacement curve.…”

Section: Resultsmentioning

confidence: 99%

“…Oligonucleotides and synthesized DNA used in this study are listed in Tables S 2 and S 3 . pDUP5-mScarlet-Ct Rac1 was generated by PCR amplification of CamScarlet with a unique 5′ AscI site and a 3′ CtRac1 followed by a unique MluI site (oligonucleotides CamScarletAscIp and yemChmCtRacMluI) and cloned into pDUP5-ADH1p-CibN-Ct Rac1 -ACT1t [ 33 ] resulting in pDUP5-ADH1p-CamScarlet-Ct Rac1 -ACT1t. The nucleotide sequence for RFP mScarlet [ 54 ] was codon optimized for C. albicans and commercially synthesized (Genscript).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were imaged as described [ 33 ] using either spinning disk confocal microscopy or wide-field fluorescence microscopy with a UPLANAPO 1.2 NA × 60 or Plan-Neofluar 0.75 NA × 40 objectives, respectively. Images were acquired at indicated times, with 0.4 or 0.5 μm z -sections (13 to 24) to capture the entire filament.…”

Section: Methodsmentioning

confidence: 99%

“…For quantitation of filament tip curvature over time, we developed another MATLAB program called TipCurve with an intuitive interface dedicated for morphological analyses along the major axis of filamentous cells. For such analyses, 3D images were converted into 2D images by sum projection, and similar to the HyphalPolarity program [ 33 ], a backbone was extracted from images over time, but also the cell contour, specifically extracting the tip first, we estimate the extremity of the backbone subtracted by the tip radius. Then, we estimate the local curvature only along the tip (either − 45° to + 45° or − 90 to 90°) from the curvilinear function f(s) extracted from cell contour: …”

Section: Methodsmentioning

confidence: 99%

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Mechanical force-induced morphology changes in a human fungal pathogen

et al. 2020

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Background The initial step of a number of human or plant fungal infections requires active penetration of host tissue. For example, active penetration of intestinal epithelia by Candida albicans is critical for dissemination from the gut into the bloodstream. However, little is known about how this fungal pathogen copes with resistive forces upon host cell invasion. Results In the present study, we have used PDMS micro-fabrication to probe the ability of filamentous C. albicans cells to penetrate and grow invasively in substrates of different stiffness. We show that there is a threshold for penetration that corresponds to a stiffness of ~ 200 kPa and that invasive growth within a stiff substrate is characterized by dramatic filament buckling, along with a stiffness-dependent decrease in extension rate. We observed a striking alteration in cell morphology, i.e., reduced cell compartment length and increased diameter during invasive growth, that is not due to depolarization of active Cdc42, but rather occurs at a substantial distance from the site of growth as a result of mechanical compression. Conclusions Our data reveal that in response to this compression, active Cdc42 levels are increased at the apex, whereas active Rho1 becomes depolarized, similar to that observed in membrane protrusions. Our results show that cell growth and morphology are altered during invasive growth, suggesting stiffness dictates the host cells that C. albicans can penetrate.

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External signal–mediated polarized growth in fungi

Bassilana

Puerner

Arkowitz

2020

Current Opinion in Cell Biology

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As the majority of fungi are non-motile, polarized growth in response to an external signal enables them to search for nutrients and mating partners, and hence is crucial for survival and proliferation. While the mechanisms underlying polarization in response to external signals has commonalities with polarization during mitotic division during budding and fission growth, the importance of diverse feedback loops regulating external signal mediated polarized growth is likely to be distinct and uniquely adapted to a dynamic environment. Here we highlight recent advances in our understanding of the mechanisms that are crucial for polarity in response to external signals in fungi, with particular focus on the roles of membrane traffic, small GTPases and lipids, as well as the interplay between cell shape and cell growth.

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Secretory Vesicle Clustering in Fungal Filamentous Cells Does Not Require Directional Growth

Cited by 17 publications

References 105 publications

The Role of Secretory Pathways in Candida albicans Pathogenesis

The Role of Secretory Pathways in Candida albicans Pathogenesis

Mechanical force-induced morphology changes in a human fungal pathogen

External signal–mediated polarized growth in fungi

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