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Article ; Online: Macropinosome formation, maturation and membrane recycling: lessons from clathrin-independent endosomal membrane systems.

Donaldson, Julie G

Philosophical transactions of the Royal Society of London. Series B, Biological sciences

2019 Volume 374, Issue 1765, Page(s) 20180148

Abstract: Macropinocytosis is a form of endocytosis that brings large fluid-filled endosomes into the cell interior. Macrophages and dendritic cells are especially active in this process, but all cells can be stimulated to initiate this remarkable form of ... ...

Abstract	Macropinocytosis is a form of endocytosis that brings large fluid-filled endosomes into the cell interior. Macrophages and dendritic cells are especially active in this process, but all cells can be stimulated to initiate this remarkable form of endocytosis. Although much is known about the membrane lipid and actin requirements for initiating macropinocytosis, less is known about the membrane that forms the macropinosome and the fate of that membrane once the macropinosome enters the cell interior. Since macropinocytosis is a specialized form of clathrin-independent endocytosis (CIE), studies of the constitutive internalization and trafficking of cargo proteins and membrane that enter cells independently of clathrin could reveal the types of membrane that form the macropinosome and the machinery that handles cargo sorting and recycling during the maturation of the macropinosome. This article is part of the Theo Murphy meeting issue 'Macropinocytosis'.
MeSH term(s)	Cell Membrane/metabolism ; Clathrin/metabolism ; Endosomes/physiology ; Pinocytosis/physiology ; Protein Transport
Chemical Substances	Clathrin
Language	English
Publishing date	2019-04-10
Publishing country	England
Document type	Journal Article
ZDB-ID	208382-6
ISSN	1471-2970 ; 0080-4622 ; 0264-3839 ; 0962-8436
ISSN (online)	1471-2970
ISSN	0080-4622 ; 0264-3839 ; 0962-8436
DOI	10.1098/rstb.2018.0148
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Article ; Online: Evaluating the Role of Galectins in Clathrin-Independent Endocytosis.

Mathew, Mohit P / Donaldson, Julie G / Hanover, John A

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2442, Page(s) 391–411

Abstract: Galectin-3 is a chimeric galectin involved in diverse intracellular and extracellular functions. Galectin-3 is synthesized in the cytoplasm and then released extracellularly by a poorly understood non-canonical secretion mechanism. As a result, it can ... ...

Abstract	Galectin-3 is a chimeric galectin involved in diverse intracellular and extracellular functions. Galectin-3 is synthesized in the cytoplasm and then released extracellularly by a poorly understood non-canonical secretion mechanism. As a result, it can play important roles both inside and outside the cell. One important extracellular role of galectin-3 is in modulating clathrin-independent endocytosis (CIE), a form of cellular internalization that is still not well understood. CIE, unlike clathrin-mediated endocytosis, has neither defined signaling sequences nor cytoplasmic machinery. As a result, extracellular interactions like the galectin-glycan interactions are thought to directly drive changes in CIE. This chapter discusses the methods designed to study the role of galectin-glycan interactions in CIE, which have provided us with insight into the functions of galectin-3 and cell surface glycans during CIE cargo internalization. These methods include media supplementation for metabolic glycoengineering, antibody internalization assays, lectin panels to assay changes in glycan patterns, exogenous galectin-3 supplementation, galectin-3 secretion assays, and in vitro assays to monitor the effect of galectins on CIE.
MeSH term(s)	Cell Membrane/metabolism ; Clathrin/metabolism ; Endocytosis/physiology ; Galectin 3/metabolism ; Methods
Chemical Substances	Clathrin ; Galectin 3
Language	English
Publishing date	2022-03-23
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2055-7_21
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Article ; Online: Nutrient-responsive O-GlcNAcylation dynamically modulates the secretion of glycan-binding protein galectin 3.

Mathew, Mohit P / Abramowitz, Lara K / Donaldson, Julie G / Hanover, John A

The Journal of biological chemistry

2022 Volume 298, Issue 3, Page(s) 101743

Abstract: Endomembrane glycosylation and cytoplasmic O-GlcNAcylation each play essential roles in nutrient sensing, and characteristic changes in glycan patterns have been described in disease states such as diabetes and cancer. These changes in glycosylation have ...

Abstract	Endomembrane glycosylation and cytoplasmic O-GlcNAcylation each play essential roles in nutrient sensing, and characteristic changes in glycan patterns have been described in disease states such as diabetes and cancer. These changes in glycosylation have important functional roles and can drive disease progression. However, little is known about the molecular mechanisms underlying how these signals are integrated and transduced into biological effects. Galectins are proteins that bind glycans and that are secreted by a poorly characterized nonclassical secretory mechanism. Once outside the cell, galectins bind to the terminal galactose residues of cell surface glycans and modulate numerous extracellular functions, such as clathrin-independent endocytosis (CIE). Originating in the cytoplasm, galectins are predicted substrates for O-GlcNAc addition and removal; and as we have shown, galectin 3 is a substrate for O-GlcNAc transferase. In this study, we also show that galectin 3 secretion is sensitive to changes in O-GlcNAc levels. We determined using immunoprecipitation and Western blotting that there is a significant difference in O-GlcNAcylation status between cytoplasmic and secreted galectin 3. We observed dramatic alterations in galectin 3 secretion in response to nutrient conditions, which were dependent on dynamic O-GlcNAcylation. Importantly, we showed that these O-GlcNAc-driven alterations in galectin 3 secretion also facilitated changes in CIE. These results indicate that dynamic O-GlcNAcylation of galectin 3 plays a role in modulating its secretion and can tune its function in transducing nutrient-sensing information coded in cell surface glycosylation into biological effects.
MeSH term(s)	Acetylglucosamine/metabolism ; Clathrin/metabolism ; Galectin 3/genetics ; Galectin 3/metabolism ; Glycosylation ; N-Acetylglucosaminyltransferases/metabolism ; Nutrients ; Polysaccharides/metabolism ; Protein Processing, Post-Translational
Chemical Substances	Clathrin ; Galectin 3 ; Polysaccharides ; N-Acetylglucosaminyltransferases (EC 2.4.1.-) ; Acetylglucosamine (V956696549)
Language	English
Publishing date	2022-02-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1016/j.jbc.2022.101743
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Article ; Online: Glycosylation and glycan interactions can serve as extracellular machinery facilitating clathrin-independent endocytosis.

Mathew, Mohit P / Donaldson, Julie G

Traffic (Copenhagen, Denmark)

2019 Volume 20, Issue 4, Page(s) 295–300

Abstract: In contrast to clathrin-mediated endocytosis (CME) which is well characterized and understood, little is known about the regulation and machinery underlying clathrin-independent endocytosis (CIE). There is also a wide variation in the requirements each ... ...

Abstract	In contrast to clathrin-mediated endocytosis (CME) which is well characterized and understood, little is known about the regulation and machinery underlying clathrin-independent endocytosis (CIE). There is also a wide variation in the requirements each individual CIE cargo has for its internalization. Recent studies have shown that CIE is affected by glycosylation and glycan interactions. We briefly review these studies and explore how these studies mesh with one another. We then discuss what this sensitivity to glycan interactions could indicate for the regulation of CIE. We address the spectrum of responses CIE has been shown to have with respect to changes in glycan interactions and attempt to reconcile disparate observations onto a shared conceptual landscape. We focus on the mechanisms by which cells can alter the glycan interactions at the plasma membrane and propose that glycosylation and glycan interactions could provide cells with a tool box with which cells can manipulate CIE. Altered glycosylation is often associated with a number of diseases and we discuss how under different disease settings, glycosylation-based modulation of CIE could play a role in disease progression.
MeSH term(s)	Animals ; Endocytosis ; Galectins/metabolism ; Glycosylation ; Humans ; Membrane Proteins/metabolism ; Polysaccharides/metabolism ; Protein Processing, Post-Translational
Chemical Substances	Galectins ; Membrane Proteins ; Polysaccharides
Language	English
Publishing date	2019-02-28
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1483852-7
ISSN	1600-0854 ; 1398-9219
ISSN (online)	1600-0854
ISSN	1398-9219
DOI	10.1111/tra.12636
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Article ; Online: Arf6, JIP3, and dynein shape and mediate macropinocytosis.

Williamson, Chad D / Donaldson, Julie G

Molecular biology of the cell

2019 Volume 30, Issue 12, Page(s) 1477–1489

Abstract: Macropinocytosis is an actin-driven form of clathrin-independent endocytosis that generates an enlarged structure, the macropinosome. Although many studies focus on signaling molecules and phosphoinositides involved in initiating macropinocytosis, the ... ...

Abstract	Macropinocytosis is an actin-driven form of clathrin-independent endocytosis that generates an enlarged structure, the macropinosome. Although many studies focus on signaling molecules and phosphoinositides involved in initiating macropinocytosis, the commitment to forming a macropinosome and the handling of that membrane have not been studied in detail. Here we show in HT1080 cells, a human fibrosarcoma cell line, a requirement for microtubules, dynein, the JIP3 microtubule motor scaffold protein, and Arf6, a JIP3 interacting protein, for the formation and inward movement of the macropinosome. While actin and myosin II also play critical roles in the formation of ruffling membrane, microtubules provide an important tract for initiation, sealing, and transport of the macropinosome through the actin- and myosin-rich lamellar region.
MeSH term(s)	ADP-Ribosylation Factors/metabolism ; Actins/metabolism ; Adaptor Proteins, Signal Transducing/metabolism ; Cell Line, Tumor ; Clathrin/metabolism ; Dyneins/metabolism ; Humans ; Microtubules/metabolism ; Models, Biological ; Mutation/genetics ; Nerve Tissue Proteins/metabolism ; Pinocytosis
Chemical Substances	Actins ; Adaptor Proteins, Signal Transducing ; Clathrin ; MAPK8IP3 protein, human ; Nerve Tissue Proteins ; Dyneins (EC 3.6.4.2) ; ADP-Ribosylation Factors (EC 3.6.5.2) ; ADP-ribosylation factor 6 (EC 3.6.5.2)
Language	English
Publishing date	2019-04-10
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E19-01-0022
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Zs.A 2853: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Immunofluorescence Staining.

Donaldson, Julie G

Current protocols in cell biology

2015 Volume 69, Page(s) 4.3.1–4.3.7

Abstract: This unit provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are exposed to the cell ... ...

Abstract	This unit provides a protocol for indirect immunofluorescence, which is a method that provides information about the locations of specific molecules and the structure of the cell. Antibody molecules for a specific target molecule are exposed to the cell or tissue being investigated. The binding of these molecules is detected by incubating the sample with a secondary antibody specific for immunoglobulin molecules and conjugated to a fluorophore. This provides both a visible signal and amplification of the signal and the results are observed with a fluorescence microscope. This unit describes the widely used and powerful technique of localization of proteins in cells by immunofluorescence. The location can be determined by double labeling with an antibody directed against a protein of known location. The technique can be used as a supplement to immunolocalization by electron microscopy and subcellular fractionation. It allows not only identification of the antigen distribution in the cell but also a survey of the dynamic aspects of protein movements in the cell-on and off membranes, into and out of the nucleus, and through membrane traffic pathways.
MeSH term(s)	Animals ; Antibodies/immunology ; Antigens/immunology ; Cells, Cultured ; Fluorescent Antibody Technique, Indirect/methods ; Fluorescent Dyes ; Humans ; Mice ; Microscopy, Fluorescence ; Organelles/metabolism ; Staining and Labeling/methods
Chemical Substances	Antibodies ; Antigens ; Fluorescent Dyes
Language	English
Publishing date	2015-12-01
Publishing country	United States
Document type	Journal Article
ISSN	1934-2616
ISSN (online)	1934-2616
DOI	10.1002/0471143030.cb0403s69
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Article ; Online: Prolonged partner separation erodes nucleus accumbens transcriptional signatures of pair bonding in male prairie voles.

Sadino, Julie M / Bradeen, Xander G / Kelly, Conor J / Brusman, Liza E / Walker, Deena M / Donaldson, Zoe R

eLife

2023 Volume 12

Abstract: The loss of a spouse is often cited as the most traumatic event in a person's life. However, for most people, the severity of grief and its maladaptive effects subside over time via an understudied adaptive process. Like humans, socially monogamous ... ...

Abstract	The loss of a spouse is often cited as the most traumatic event in a person's life. However, for most people, the severity of grief and its maladaptive effects subside over time via an understudied adaptive process. Like humans, socially monogamous prairie voles (
MeSH term(s)	Animals ; Humans ; Male ; Nucleus Accumbens ; Grassland ; Pair Bond ; Arvicolinae/genetics ; DNA-Binding Proteins ; Social Behavior
Chemical Substances	DNA-Binding Proteins
Language	English
Publishing date	2023-02-28
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.80517
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Article ; Online: Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin-glycan interactions in clathrin-independent endocytosis.

Mathew, Mohit P / Donaldson, Julie G

The Journal of biological chemistry

2018 Volume 293, Issue 19, Page(s) 7222–7237

Abstract: Clathrin-independent endocytosis (CIE) is a form of endocytosis that lacks a defined cytoplasmic machinery. Here, we asked whether glycan interactions, acting from the outside, could be a part of that endocytic machinery. We show that the perturbation of ...

Abstract	Clathrin-independent endocytosis (CIE) is a form of endocytosis that lacks a defined cytoplasmic machinery. Here, we asked whether glycan interactions, acting from the outside, could be a part of that endocytic machinery. We show that the perturbation of global cellular patterns of protein glycosylation by modulation of metabolic flux affects CIE. Interestingly, these changes in glycosylation had cargo-specific effects. For example, in HeLa cells, GlcNAc treatment, which increases glycan branching, increased major histocompatibility complex class I (MHCI) internalization but inhibited CIE of the glycoprotein CD59 molecule (CD59). The effects of knocking down the expression of galectin 3, a carbohydrate-binding protein and an important player in galectin-glycan interactions, were also cargo-specific and stimulated CD59 uptake. By contrast, inhibition of all galectin-glycan interactions by lactose inhibited CIE of both MHCI and CD59. None of these treatments affected clathrin-mediated endocytosis, implying that glycosylation changes specifically affect CIE. We also found that the galectin lattice tailors membrane fluidity and cell spreading. Furthermore, changes in membrane dynamics mediated by the galectin lattice affected macropinocytosis, an altered form of CIE, in HT1080 cells. Our results suggest that glycans play an important and nuanced role in CIE, with each cargo being affected uniquely by alterations in galectin and glycan profiles and their interactions. We conclude that galectin-driven effects exist on a continuum from stimulatory to inhibitory, with distinct CIE cargo proteins having unique response landscapes and with different cell types starting at different positions on these conceptual landscapes.
MeSH term(s)	Acetylglucosamine/pharmacology ; CD59 Antigens/metabolism ; Cell Membrane/drug effects ; Cell Movement/physiology ; Clathrin/physiology ; Culture Media ; Endocytosis/physiology ; Galectin 3/genetics ; Galectin 3/metabolism ; Galectin 3/pharmacology ; Gene Knockdown Techniques ; Glycosylation ; HeLa Cells ; Histocompatibility Antigens Class I/metabolism ; Humans ; Lactose/pharmacology ; Membrane Fluidity/physiology ; Pinocytosis/physiology ; Polysaccharides/metabolism ; Protein Transport/physiology
Chemical Substances	CD59 Antigens ; Clathrin ; Culture Media ; Galectin 3 ; Histocompatibility Antigens Class I ; Polysaccharides ; CD59 protein, human (101754-01-2) ; Lactose (J2B2A4N98G) ; Acetylglucosamine (V956696549)
Language	English
Publishing date	2018-03-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.RA118.001802
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Article ; Online: Prolonged partner separation erodes nucleus accumbens transcriptional signatures of pair bonding in male prairie voles

Julie M Sadino / Xander G Bradeen / Conor J Kelly / Liza E Brusman / Deena M Walker / Zoe R Donaldson

eLife, Vol

2023 Volume 12

Abstract: The loss of a spouse is often cited as the most traumatic event in a person’s life. However, for most people, the severity of grief and its maladaptive effects subside over time via an understudied adaptive process. Like humans, socially monogamous ... ...

Abstract	The loss of a spouse is often cited as the most traumatic event in a person’s life. However, for most people, the severity of grief and its maladaptive effects subside over time via an understudied adaptive process. Like humans, socially monogamous prairie voles (Microtus ochrogaster) form opposite-sex pair bonds, and upon partner separation, show stress phenotypes that diminish over time. We test the hypothesis that extended partner separation diminishes pair bond-associated behaviors and causes pair bond transcriptional signatures to erode. Opposite-sex or same-sex paired males were cohoused for 2 weeks and then either remained paired or were separated for 48 hours or 4 weeks before collecting fresh nucleus accumbens tissue for RNAseq. In a separate cohort, we assessed partner-directed affiliation at these time points. We found that these behaviors persist despite prolonged separation in both same-sex and opposite-sex paired voles. Opposite-sex pair bonding led to changes in accumbal transcription that were stably maintained while animals remained paired but eroded following prolonged partner separation. Eroded genes are associated with gliogenesis and myelination, suggesting a previously undescribed role for glia in pair bonding and loss. Further, we pioneered neuron-specific translating ribosomal affinity purification in voles. Neuronally enriched transcriptional changes revealed dopaminergic-, mitochondrial-, and steroid hormone signaling-associated gene clusters sensitive to acute pair bond disruption and loss adaptation. Our results suggest that partner separation erodes transcriptomic signatures of pair bonding despite core behavioral features of the bond remaining intact, revealing potential molecular processes priming a vole to be able to form a new bond.
Keywords	prairie vole ; nucleus accumbens ; loss ; pair bond ; Medicine ; R ; Science ; Q ; Biology (General) ; QH301-705.5
Subject code	590
Language	English
Publishing date	2023-02-01T00:00:00Z
Publisher	eLife Sciences Publications Ltd
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Hyperbaric oxygen as a model of lens aging in the bovine lens: The effects on lens biochemistry, physiology and optics.

Lim, Julie C / Grey, Angus C / Vaghefi, Ehsan / Nye-Wood, Mitchell G / Donaldson, Paul J

Experimental eye research

2021 Volume 212, Page(s) 108790

Abstract: Age related nuclear (ARN) cataracts in humans take years to form and so experimental models have been developed to mimic the process in animals as a means of better understanding the etiology of nuclear cataracts in humans. A major limitation with these ... ...

Abstract	Age related nuclear (ARN) cataracts in humans take years to form and so experimental models have been developed to mimic the process in animals as a means of better understanding the etiology of nuclear cataracts in humans. A major limitation with these animal models is that many of the biochemical and physiological changes are not typical of that seen in human ARN cataract. In this review, we highlight the work of Frank Giblin and colleagues who established an in vivo animal model that replicates many of the changes observed in human ARN cataract. This model involves exposing aged guinea pigs to hyperbaric oxygen (HBO), which by causing the depletion of the antioxidant glutathione (GSH) specifically in the lens nucleus, produces oxidative changes to nuclear proteins, nuclear light scattering and a myopic shift in lens power that mimics the change that often precedes cataract development in humans. However, this model involves multiple HBO treatments per week, with sometimes up to a total of 100 treatments, spanning up to eight months, which is both costly and time consuming. To address these issues, Giblin developed an in vitro model that used rabbit lenses exposed to HBO for several hours which was subsequently shown to replicate many of the changes observed in human ARN cataract. These experiments suggest that HBO treatment of in vitro animal lenses may serve as a more economical and efficient model to study the development of cataract. Inspired by these experiments, we investigated whether exposure of young bovine lenses to HBO for 15 h could also serve as a suitable acute model of ARN cataract. We found that while this model is able to exhibit some of the biochemical and physiological changes associated with ARN cataract, the decrease in lens power we observed was more characteristic of the hyperopic shift in refraction associated with ageing. Future work will investigate whether HBO treatment to age the bovine lens in combination with an oxidative stressor such as UV light will induce refractive changes more closely associated with human ARN cataract. This will be important as developing an animal model that replicates the changes to lens biochemistry, physiology and optics observed in human ARN cataracts is urgently required to facilitate the identification and testing of anti-cataract therapies that are effective in humans.
MeSH term(s)	Aging ; Animals ; Cataract/metabolism ; Cataract/physiopathology ; Cattle ; Humans ; Hyperbaric Oxygenation/methods ; Lens, Crystalline/chemistry ; Lens, Crystalline/diagnostic imaging ; Lens, Crystalline/physiology ; Optics and Photonics ; Slit Lamp Microscopy
Language	English
Publishing date	2021-10-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	80122-7
ISSN	1096-0007 ; 0014-4835
ISSN (online)	1096-0007
ISSN	0014-4835
DOI	10.1016/j.exer.2021.108790
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