CP39, CP75 and CP91 are major structural components of the Dictyostelium centrosome’s core structure

Meyer, Irene; Peter, Tatjana; Batsios, Petros; Kuhnert, Oliver; Krüger‐Genge, Anne; Camurça, Carl; Gräf, Ralph

doi:10.1016/j.ejcb.2017.01.004

Cited by 13 publications

(31 citation statements)

References 47 publications

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“…Transformation of Dictyostelium amoebae by electroporation, SDS electrophoresis and Western blotting were carried out according to standard procedures [ 25 , 29 ]. Centrosomes or nuclei with attached centrosomes and were isolated as reported earlier [ 30 ]. BioID assays were performed as described recently [ 20 , 30 ].…”

Section: Methodsmentioning

confidence: 99%

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CDK5RAP2 Is an Essential Scaffolding Protein of the Corona of the Dictyostelium Centrosome

Pitzen

Askarzada

Gräf

et al. 2018

Cells

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Dictyostelium centrosomes consist of a nucleus-associated cylindrical, three-layered core structure surrounded by a corona consisting of microtubule-nucleation complexes embedded in a scaffold of large coiled-coil proteins. One of them is the conserved CDK5RAP2 protein. Here we focus on the role of Dictyostelium CDK5RAP2 for maintenance of centrosome integrity, its interaction partners and its dynamic behavior during interphase and mitosis. GFP-CDK5RAP2 is present at the centrosome during the entire cell cycle except from a short period during prophase, correlating with the normal dissociation of the corona at this stage. RNAi depletion of CDK5RAP2 results in complete disorganization of centrosomes and microtubules suggesting that CDK5RAP2 is required for organization of the corona and its association to the core structure. This is in line with the observation that overexpressed GFP-CDK5RAP2 elicited supernumerary cytosolic MTOCs. The phenotype of CDK5RAP2 depletion was very reminiscent of that observed upon depletion of CP148, another scaffolding protein of the corona. BioID interaction assays revealed an interaction of CDK5RAP2 not only with the corona markers CP148, γ-tubulin, and CP248, but also with the core components Cep192, CP75, and CP91. Furthermore, protein localization studies in both depletion strains revealed that CP148 and CDK5RAP2 cooperate in corona organization.

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Section: Methodsmentioning

confidence: 99%

“…Centrosomes or nuclei with attached centrosomes and were isolated as reported earlier [ 30 ]. BioID assays were performed as described recently [ 20 , 30 ].…”

Section: Methodsmentioning

confidence: 99%

CDK5RAP2 Is an Essential Scaffolding Protein of the Corona of the Dictyostelium Centrosome

Pitzen

Askarzada

Gräf

et al. 2018

Cells

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“…Concerning cell motility, an uninterrupted series of studies in the last 50 years has led to the identification and characterization of the acto-myosin cytoskeleton underlying changes in cell shape and cell motility processes, with many cytoskeletal proteins first identified and/or characterized in Dictyostelium, such as coronin, the actin nucleator SCAR, the 34-kDa actin-crosslinking protein, myosin I and II, and formins (see Bozzaro, 2013 for references). The dynamic structure of the cell cortex, the actin cytoskeleton-nuclear membrane interactions, nucleus/nucleolus and the microtubule cytoskeleton, and their regulation by small GTPases have been the subject of several studies also in recent years (Nichols et al, 2015;Gräf et al, 2015;Rivero and Xiong, 2016;Meyer et al, 2017;Pitzen et al, 2018). Recently, the role of formins in regulating the functional integrity of the cell cortex has been investigated in detail (Junemann et al, 2016), and it has been shown that the HSBP1 protein, which regulates WASH complex assembly at centrosomes, is required for development of focal adhesion and cell polarity in Dictyostelium as well as in tumour cells (Visweshwaran et al, 2018).…”

Section: Past and Present Of A Model Organismmentioning

confidence: 99%

The past, present and future of Dictyostelium as a model system

Bozzaro¹

2019

Int. J. Dev. Biol.

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The social amoeba Dictyostelium discoideum has been a preferred model organism during the last 50 years, particularly for the study of cell motility and chemotaxis, phagocytosis and macropinocytosis, intercellular adhesion, pattern formation, caspase-independent cell death and more recently autophagy and social evolution. Being a soil amoeba and professional phagocyte, thus exposed to a variety of potential pathogens, D. discoideum has also proven to be a powerful genetic and cellular model for investigating host-pathogen interactions and microbial infections. The finding that the Dictyostelium genome harbours several homologs of human genes responsible for a variety of diseases has stimulated their analysis, providing new insights into the mechanism of action of the encoded proteins and in some cases into the defect underlying the disease. Recent technological developments have covered the genetic gap between mammals and non-mammalian model organisms, challenging the modelling role of the latter. Is there a future for Dictyostelium discoideum as a model organism? Peculiarities and advantages of Dictyostelium discoideum as model organismAmong the non-mammalian model organisms, Dictyostelium discoideum (in the following Dictyostelium) is unique, due to cell division and development being totally uncoupled, and because of the transition from a unicellular to a multicellular stage during the life cycle. Growing cells proliferate by binary fission but do not differentiate, whereas starving cells undergo multicellular development and differentiation without dividing and without any need for external nutrients. Thus growth and development can be studied separately, and several non-lethal mutants can be isolated

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“…The centrosome inserts into the nuclear envelope at the G2/M transition, duplicates and forms the two spindle poles. Failure of this insertion process leads to mitotic delays and supernumerary centrosome formation (Leo et al, 2012;Meyer et al, 2017). Centrosome insertion certainly requires close proximity to the nuclear envelope, but also deformation of the nuclear envelope in order to form a fenestra which ultimately harbours the duplicating centrosome.…”

Section: The Dynamics Of the Nuclear Envelope During Mitosismentioning

confidence: 99%

“…The mechanism how this fenestra forms is unknown. However, a recent study has revealed that the centrosomal protein CP75 is necessary for permeabilization of the nuclear envelope at the G2/M transition (Meyer et al, 2017). Transmission EM observations further suggest that the nuclear envelope is a contiguous structure during nuclear division, without further fenestrations until late telophase, when the nuclear envelope bridge between the two separating daughter nuclei ruptures resulting in two distinct fenestrae, one at each daughter nucleus, that are penetrated by the central spindle (McIntosh et al, 1985).…”

Section: The Dynamics Of the Nuclear Envelope During Mitosismentioning

confidence: 99%

Nuclear envelope organization in Dictyostelium discoideum

Batsios¹,

Gräf²,

Koonce³

et al. 2019

Int. J. Dev. Biol.

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The nuclear envelope consists of the outer and the inner nuclear membrane, the nuclear lamina and the nuclear pore complexes, which regulate nuclear import and export. The major constituent of the nuclear lamina of Dictyostelium is the lamin NE81. It can form filaments like B-type lamins and it interacts with Sun1, as well as with the LEM/HeH-family protein Src1. Sun1 and Src1 are nuclear envelope transmembrane proteins involved in the centrosome-nucleus connection and nuclear envelope stability at the nucleolar regions, respectively. In conjunction with a KASH-domain protein, Sun1 usually forms a so-called LINC complex. Two proteins with functions reminiscent of KASH-domain proteins at the outer nuclear membrane of Dictyostelium are known; interaptin which serves as an actin connector and the kinesin Kif9 which plays a role in the microtubule-centrosome connector. However, both of these lack the conserved KASH-domain. The link of the centrosome to the nuclear envelope is essential for the insertion of the centrosome into the nuclear envelope and the appropriate spindle formation. Moreover, centrosome insertion is involved in permeabilization of the mitotic nucleus, which ensures access of tubulin dimers and spindle assembly factors. Our recent progress in identifying key molecular players at the nuclear envelope of Dictyostelium promises further insights into the mechanisms of nuclear envelope dynamics. , and build upon the material for any purpose, even commercially), providing you give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use. Printed in Spain Abbreviations used in this paper: BAF, barrier to autointegration factor; BioID, proximity-dependent biotin identification; CDK1, cyclin-dependent kinase 1; cryo-ET, cryo-electron tomography; DAPI, 4′,6-diamidin-2-phenylindol; EM, electron microscopy; ER, endoplasmic reticulum; ExM, expansion microscopy; feSEM, field-emission scanning electron microscopy; FRAP, fluorescence recovery after photobleaching; GFP, green fluorescent protein; HeH, helix-extension-helix; IF, intermediate filament; INM, inner nuclear membrane; KASH, Klarsicht, ANC-1, Syne Homology; LBR, lamin B receptor; LECA, last eukaryotic common ancestor; LEM, lamina-associated polypeptide 2 (LAP2), Emerin, and MAN1; LINC, linker of the nucleoskeleton and the cytoskeleton; MCAK, mitotic centromere-associated kinesin; MEF, mouse embryonic fibroblast; NET, nuclear envelope transmembrane protein; NLS, nuclear localization sequence; NPC, nuclear pore complex; ONM, outer nuclear membrane; STED, stimulated emission depletion. import and export. On its nuclear side, the INM is associated with a fibrous, so-called nuclear lamina. Among the NETs, the Sun-domain proteins in the INM (e.g. Sun1) and KASH-domain proteins in the ONM (e.g. nesprin) have to be highlighted, as they form the so-called LINC complex, which spans the whole nuclear envelope (Lee and Burke, 2018...

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CP39, CP75 and CP91 are major structural components of the Dictyostelium centrosome’s core structure

Cited by 13 publications

References 47 publications

CDK5RAP2 Is an Essential Scaffolding Protein of the Corona of the Dictyostelium Centrosome

CDK5RAP2 Is an Essential Scaffolding Protein of the Corona of the Dictyostelium Centrosome

The past, present and future of Dictyostelium as a model system

Nuclear envelope organization in Dictyostelium discoideum

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