Localization of Centrins in the Hypotrich Ciliate Paraurostyla weissei

Lemullois, Michel; Fryd-Versavel, G.; Fleury-Aubusson, Anne

doi:10.1078/1434461041844178

Cited by 11 publications

(16 citation statements)

References 45 publications

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“…The same localisation has been observed in another ciliate, Paraurostyla (Lemullois et al, 2004). We show here that, in addition to this proximal localisation, some PtCen3p is also detected distally, within the basal body at the transition zone.…”

Section: Ptcen3p Is Required For Basal Body Docking Of Fully Mature Bsupporting

confidence: 58%

FOR20, a conserved centrosomal protein, is required for assembly of the transition zone and basal body docking at the cell surface

Aubusson-Fleury

Lemullois

Loubresse

et al. 2012

Journal of Cell Science

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SummaryWithin the FOP family of centrosomal proteins, the conserved FOR20 protein has been implicated in the control of primary cilium assembly in human cells. To ascertain its role in ciliogenesis, we have investigated the function of its ortholog, PtFOR20p, in the multiciliated unicellular organism Paramecium. Using combined functional and cytological analyses, we found that PtFOR20p specifically localises at basal bodies and is required to build the transition zone, a prerequisite to their maturation and docking at the cell surface and hence to ciliogenesis. We also found that PtCen2p (one of the two basal body specific centrins, an ortholog of HsCen2) is required to recruit PtFOR20p at the developing basal body and to control its length. By contrast, the other basal-body-specific centrin PtCen3p is not needed for assembly of the transition zone, but is required downstream, for basal body docking. Comparison of the structural defects induced by depletion of PtFOR20p, PtCen2p or PtCen3p, respectively, illustrates the dual role of the transition zone in the biogenesis of the basal body and in cilium assembly. The multiple potential roles of the transition zone during basal body biogenesis and the evolutionary conserved function of the FOP proteins in microtubule membrane interactions are discussed.

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Section: Ptcen3p Is Required For Basal Body Docking Of Fully Mature Bsupporting

confidence: 58%

FOR20, a conserved centrosomal protein, is required for assembly of the transition zone and basal body docking at the cell surface

Aubusson-Fleury

Lemullois

Loubresse

et al. 2012

Journal of Cell Science

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“…Extraction was conducted using the method described by Lemullois et al (2004). Cells were permeabilized in the cytoskeleton-stabilizing buffer PHEM containing 1% Triton X100 and supplemented with protease inhibitors (leupeptin 2 mM; pepstatin 1.5 mM; aprotinin 0.15 mM; benzamidin 10 mM) for 10 minutes on ice.…”

Section: Preparation Of Protein Extractsmentioning

confidence: 99%

“…However, not much is known about the functions and structures of these proteins in hypotrich ciliates (Liang et al 1996;Lemullois et al 2004;Marziale et al 2008). Here we analyzed the subcellular localization and function of a-, g-, and d-tubulins in Stylonychia pustulata.…”

Section: Introductionmentioning

confidence: 99%

“…Hypotrich ciliates contain thousands of cilia and associated centriolar-like basal bodies, which are rapidly assembled, making the ciliated protozoa excellent models for investigating the role of cytoskeletal and basal bodyÁassociated proteins (Lemullois et al 2004). However, not much is known about the functions and structures of these proteins in hypotrich ciliates (Liang et al 1996;Lemullois et al 2004;Marziale et al 2008).…”

Section: Introductionmentioning

confidence: 99%

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Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

Yin

Sheng

Gao

et al. 2013

Animal Cells and Systems

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Localization of α-, γ-, and δ-tubulin in the hypotrich ciliate Stylonychiapustulata (Hyportrichida, Ciliophora), Animal Cells and Systems, 17:4, 259-268, DOI: 10.1080/19768354.2013.828654 To link to this article: https://doi.org/10. 1080/19768354.2013 Ciliature microtubular organelles, which organize into ciliary units, are highly complex microtubular systems. To investigate the formation of ciliature microtubular organelles and the role that different tubulins play in this process, we examined the distribution and function of a-, g-, and d-tubulins in the hypotrich ciliate Stylonychia pustulata. Immunofluorescence analysis demonstrated that a-tubulin localized to all microtubular organelles, while g-and dtubulins colocalized at basal bodies. Immunogold labeling revealed that g-and d-tubulins colocalize to the proximal part of basal bodies, suggesting that g-and d-tubulins probably interact and may function together. In addition, the permanent localization of g-tubulin to the adoral zone of membranelles suggests that g-tubulin play a role in the maintenance of oral ciliature base-associated microtubules, but does not play a role in the biogenesis of these ciliature. We observe that d-tubulin is restricted to the basal body and the connective fibers of the basal body, suggesting this protein may be required for the extension and stability of the microtubules in these structures. Dramatically different concentrations of these three tubulins were observed in the cytoskeleton and the insoluble and soluble fractions of the cell, suggesting that these highly related proteins perform different functions in S. pustulata.

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“…In the ciliate Paramecium, three centrin genes have been characterized to date (38) and in another ciliate (Paraurostyla) evidence has been presented for differential localization of two centrin isotypes (36).…”

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confidence: 99%

Centrin Scaffold in Chlamydomonas reinhardtii Revealed by Immunoelectron Microscopy

Geimer

Melkonian

2005

Eukaryot Cell

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In the flagellate green alga Chlamydomonas reinhardtii the Ca 2؉ -binding EF-hand protein centrin is encoded by a single-copy gene. Previous studies have localized the protein to four distinct structures in the flagellar apparatus: the nucleus-basal body connector, the distal connecting fiber, the flagellar transitional region, and the axoneme. To explain the disjunctive distribution of centrin, the interaction of centrin with as yet unknown specific centrin-binding proteins has been implied. Here, we demonstrate using serial section postembedding immunoelectron microscopy of isolated cytoskeletons that centrin is located in additional structures (transitional fibers and basal body lumen) and that the centrin-containing structures of the basal apparatus are likely part of a continuous filamentous scaffold that extends from the nucleus to the flagellar bases. In addition, we show that centrin is located in the distal lumen of the basal body in a rotationally asymmetric structure, the V-shaped filament system. This novel centrin-containing structure has also been detected near the distal end of the probasal bodies. Taken together, these results suggest a role for a rotationally asymmetric centrin "seed" in the growth and development of the centrin scaffold following replication of the basal apparatus.Centrin (also known as caltractin) is a member of the highly conserved superfamily of calcium-binding EF-hand proteins and occurs in all eukaryotic cells, being mostly associated with centrosomes (58, 64). It was first discovered in flagellate green algae, where it is located in the basal apparatus (the part of the cytoskeleton anchoring the flagella) and involved in the contraction of calcium-sensitive filaments (42, 60) which, in general, play an important role in the dynamic behavior of centrosomes by controlling the position and orientation of centrosomal structures (58, 64). Analysis of centrin mutants and RNA interference strains deficient in centrin in the green algal model system Chlamydomonas reinhardtii suggested that centrin is essential for basal body duplication and segregation during cell division (28,39,73), functions also deduced for centrin homologues in vertebrates (46, 62) and yeast spp. (3,50,69). More recently, novel centrin functions have been discovered such as involvement in recombinational DNA repair through modulation of the nucleotide excision repair pathway in humans and plants (2, 47), a role in the nuclear mRNA export machinery in yeast (11), regulation of calcium-dependent ciliary activities in ciliates (14, 15), and signal transduction in vertebrate photoreceptors (53). How centrin exerts these different functions at the molecular level is, however, still unknown although considerable progress has recently been made through in vitro studies of centrin and its interaction with target peptides (8,19,20,40,75).In some organisms, the functional diversity of centrin may be related to the presence of different centrin isotypes, e.g., four centrin proteins (centrin1p to centrin4p), which are d...

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Localization of Centrins in the Hypotrich Ciliate Paraurostyla weissei

Cited by 11 publications

References 45 publications

FOR20, a conserved centrosomal protein, is required for assembly of the transition zone and basal body docking at the cell surface

FOR20, a conserved centrosomal protein, is required for assembly of the transition zone and basal body docking at the cell surface

Localization of α-, γ-, and δ-tubulin in the hypotrich ciliateStylonychia pustulata(Hyportrichida, Ciliophora)

Centrin Scaffold in Chlamydomonas reinhardtii Revealed by Immunoelectron Microscopy

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