Synthesis and turnover of cell body-agglutinin as a pool of flagellar surface-agglutinin inChlamydomonas reinhardii gamete

Saito, Tatsuaki; Tsubo, Yoshihiro; Matsuda, Yoshihisa

doi:10.1007/bf00693391

Cited by 45 publications

(36 citation statements)

References 16 publications

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“…We showed that as cells adhere to each other they continuously inactivate the adhesiveness of each other's flagella (42,44). Replacement of these lost adhesive sites requires protein synthesis (36,37,42) and protein glycosylation (29, 39, 48); Cooper et al showed that hydroxylation of proline was also required for replacement of these lost adhesion sites (10). Similar results have been shown for Chlamydomonas eugametos (32,33,45,(47)(48)(49).…”

mentioning

confidence: 73%

“…Recently Goodenough, Heuser, Adair, and co-workers identified high molecular mass glycoproteins rich in hydroxyproline that are involved in the adhesive interaction (1,2,4,9,10,16). Saito and Matsuda (35,37) obtained similar results using different purification methods, and van den Ende's laboratory has identified similar high molecular mass agglutinin molecules in C. eugametos (20,33,45).…”

mentioning

confidence: 77%

“…Flagella isolated by use of the sucrose-pH shock method of Witman et al (50) as previously described (37), were resuspended in the n-octylglucoside (OG) extraction buffer described by Adair et al (4) that contained 40 mM OG, 0.1 mM dithiothreitol, 0.1 mM EGTA and 10 mM Tris, pH 7.3. Routinely, flagella isolated from 16 liters of cells were resuspended in about 2 ml of OG extraction buffer.…”

Section: Preparation Of Detergent Extracts Of Flagellamentioning

confidence: 99%

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A monoclonal antibody that blocks adhesion of Chlamydomonas mt+ gametes.

Snell¹,

Clausell

Perillo³

et al. 1986

The Journal of Cell Biology

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Abstract. During the mating reaction (fertilization) in the biflagellated alga, Chlamydomonas reinhardtii, mt* and mt-gametes adhere to each other via their flagella and subsequently fuse to form quadriflagellated zygotes. In the studies reported here, we describe a monoclonal antibody directed against an mt + flagellar surface molecule. The antibody blocks the adhesiveness of mt + gametes, isolated mt ÷ flagella, and detergent extracts thereof. It has no effect on mt-gametes. Cyanogen bromide-activated Sepharose beads derivatized with the antibody bind only mt + gametes; mt-gametes and mt ÷ and mt-vegetative cells are unreactive with the derivatized beads. The interaction of mt + gametes with the beads is dynamic and cells continuously bind, detach, and rebind to the beads. Surprisingly, antibody-derivatized beads that have been incubated with mt + gametes acquire the ability to bind mr-gametes. Moreover, extraction of the preincubated beads with detergents releases active mt + adhesion molecules. The evidence suggests that binding of the antibody to the flagellar surface adhesion molecules causes their release from the flagellar surface, possibly mimicking the normal mechanism of flagellar de-adhesion.W HEN gametes of opposite mating types of the biflagellated alga Chlamydomonas reinhardtii are mixed together, they adhere to each other via their flagella and form large clumps of aggregating cells. Within these clumps, pairs of cells form that eventually fuse to become quadriflagellated zygotes. Several laboratories have been interested in the initial adhesive interaction. Recently Goodenough, Heuser, Adair, and co-workers identified high molecular mass glycoproteins rich in hydroxyproline that are involved in the adhesive interaction (1, 2, 4, 9, 10, 16). Saito and Matsuda (35, 37) obtained similar results using different purification methods, and van den Ende's laboratory has identified similar high molecular mass agglutinin molecules in C. eugametos (20,33, 45).Our laboratory has been interested in the molecular details of the adhesive interaction as well as a specific de-adhesion mechanism that we have characterized (39,(40)(41)(42)44). We showed that as cells adhere to each other they continuously inactivate the adhesiveness of each other's flagella (42, 44). Replacement of these lost adhesive sites requires protein synthesis (36, 37, 42) and protein glycosylation (29, 39, 48); Cooper et al. showed that hydroxylation of proline was also required for replacement of these lost adhesion sites (10). Similar results have been shown for Chlamydomonas eugametos (32,33, 45,(47)(48)(49). To more fully study this adhesion-induced loss of flagellar adhesion sites and to identify molecules responsible for adhesion we have begun to use an immunological approach. In earlier unpublished studies we had attempted to prepare rabbit polyclonal antibodies against flagella or flagellar surface fractions that would specifically block adhesiveness of mating type plus (mr+) 1 or mating type minus (mr-) gametes. To date, all of th...

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confidence: 73%

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confidence: 77%

Section: Preparation Of Detergent Extracts Of Flagellamentioning

confidence: 99%

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A monoclonal antibody that blocks adhesion of Chlamydomonas mt+ gametes.

Snell¹,

Clausell

Perillo³

et al. 1986

The Journal of Cell Biology

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“…Agglutinins are Iocalized on ff agelIa (ff agellar agglutinin) and the cell body (cell body agglutinin) (Saito et al, 1985;Musgrave et al, 1986;Hunnicutt et al, 1990). After an incubation of gametes with cycloheximide for 6 h, active flagellar agglutinin disappeared, whereas a significant amount of cell body agglutinin remained (Fig.…”

Section: Control Of Mating Competence By Lightmentioning

confidence: 97%

Characterization of Blue Light Signal Transduction Chains That Control Development and Maintenance of Sexual Competence in Chlamydomonas reinhardtii

1997

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Blue light induces the differentiation of Chlamydomonas reinhardtii pregametes to gametes. The light-induced conversion of pregametes to gametes is protein synthesis dependent and proceeds only after a lag phase. Upon incubation in the dark, gametes lost their mating ability, resulting in dark-inactivated gametes. Reillumination rapidly restored mating competence and this was shown to be independent of protein synthesis. Apparently, differentiation and maintenance of gametic competence are both regulated by light. Whether one or two light-activated signal pathways are involved was investigated using pharmacological compounds that affect signal transduction. Compounds that affected pregamete-togamete conversion affected the expression of a gamete-specific gene in a similar fashion. Other drugs affected only dark-inactivated gametes, suggesting that reactivating gametes requires a separate signaling pathway. Combined treatments provided evidence for the consecutive action of a phosphatase and a protein kinase C-like kinase in the light-induced reactivation process.

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“…These and other changes, e.g. the appearance of sexual agglutinins (1,11,19) and the synthesis of gametic autolysin (12,16) suggest that gametic differentiation is based on changes in the pattern of gene expression. Characteristic differences in the rates of synthesis of individual proteins (15) and the accumulation of mRNA species specific for gametes (von Gromoff, C.F.…”

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confidence: 99%

Gametic Differentiation of Chlamydomonas reinhardtii

Beck

Acker²

1992

Plant Physiol.

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Gametic differentiation of the unicellular green alga Chiamydomonas reinhardtii proceeds in two steps controlled by the extrinsic signals nitrogen deficiency and light. Nitrogen deprivation induces the differentiation of vegetative cells to sexually immature pregametes. A light signal is required to convert the pregametes to gametes. Both signals are also required for the maintenance of mating competence. Two converging signal transduction chains are proposed to control gamete formation. For the differentiation of pregametes to gametes, a fluence ratedependent reaction, requiring continuous irradiation, is suggested by photobiological experiments.By gametic differentiation, Chlamydomonas reinhardtii vegetative cells are transformed into mating-competent cells. This differentiation has been shown to be associated with changes in the cell's biochemistry and subcellular morphology. The most prominent changes at the molecular level have been demonstrated for the ribosomes. Cytoplasmic and chloroplast ribosomes decrease in number (7,8,13) and new ribosomes appear that exhibit decreased translational accuracy (2), altered susceptibility to antibiotics (9), and ribosomal proteins with different electrophoretic mobilities (9). Morphologically, the acquisition of two new organelles, a mating structure and a special type of Golgi-derived vesicle are most prominent (3, 7). These and other changes, e.g. the appearance of sexual agglutinins (1,11,19) and the synthesis of gametic autolysin (12,16) suggest that gametic differentiation is based on changes in the pattern of gene expression. Characteristic differences in the rates of synthesis of individual proteins (15)

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Synthesis and turnover of cell body-agglutinin as a pool of flagellar surface-agglutinin inChlamydomonas reinhardii gamete

Cited by 45 publications

References 16 publications

A monoclonal antibody that blocks adhesion of Chlamydomonas mt+ gametes.

A monoclonal antibody that blocks adhesion of Chlamydomonas mt+ gametes.

Characterization of Blue Light Signal Transduction Chains That Control Development and Maintenance of Sexual Competence in Chlamydomonas reinhardtii

Gametic Differentiation of Chlamydomonas reinhardtii

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