Chapter 15 Probing the Mechanisms of Mitosis with Dictyostelium discoideum

Roos, Urs-Peter

doi:10.1016/s0091-679x(08)61650-7

Cited by 19 publications

(11 citation statements)

References 59 publications

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“…Coverslip specimens were essentially prepared as described Roos, 1994, 1995;Roos, 1987;Roos and Guhl, 1996). Coverslips cleaned in 70% ethanol and placed in a humid chamber were seeded with a drop from a liquid culture.…”

Section: Cytochemistrymentioning

confidence: 99%

“…The secondary antibody was a DTAFlabeled goat anti-mouse IgG ϩ IgM (Jackson ImmunoResearch Laboratories Inc., Dianova, Hamburg, Germany). Cells fixed with FA/GA were treated twice for 5 min with 0.1% NaBH 4 before the primary antibody was applied and the preparations were processed further as described (Roos, 1987). We also double-labeled amoebae with rhodamine-phalloidin and the anti-actin antibody to compare the immunofluorescence patterns of total actin and F-actin.…”

Section: Cytochemistrymentioning

confidence: 99%

“…Amoebae of D. discoideum NC-4 were grown to log phase in a medium modified from the formula of Sussman (Sussman, 1961) (see Roos, 1987) and strain Ax-2 was grown in HL-5 medium (Cocucci and Sussman, 1970). A. rosea and P. mycophaga were grown in the medium of Spiegel (Guhl and Roos, 1995;Roos and Guhl, 1996;Spiegel, 1982).…”

mentioning

confidence: 99%

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The Actomyosin Cytoskeleton of Amoebae of the Cellular Slime MoldsAcrasis roseaandProtostelium mycophaga: Structure, Biochemical Properties, and Function

Hellstén

Roos

1998

Fungal Genetics and Biology

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Section: Cytochemistrymentioning

confidence: 99%

Section: Cytochemistrymentioning

confidence: 99%

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The Actomyosin Cytoskeleton of Amoebae of the Cellular Slime MoldsAcrasis roseaandProtostelium mycophaga: Structure, Biochemical Properties, and Function

Hellstén

Roos

1998

Fungal Genetics and Biology

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“…Microtubules were depolymerized with nocodazole, the most effective reagent in Dictyostelium (42). Model Hirano bodies in Dictyostelium are highly enriched for myosin II, but not myosin I (16).…”

Section: Resultsmentioning

confidence: 99%

Requirements for Hirano Body Formation

et al. 2014

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“…Dictyostelium amoebae grown in suspension to a final density of 1-5 x 105 were allowed to attach to polylysine-coated coverslips and fixed with 3.7% (vol/vol) formaldehyde in a cytoskeleton buffer (16), according to the two-step fixation protocol of Roos (21). HeLa and EMF cells were cultured directly on coverslips in Dulbecco's minimal Eagle's medium or F-10 medium, washed two times with phosphate-buffered saline, and then fixed for 10 min with 4% formaldehyde in 100 mM Pipes (pH 6.9)/5 mM EGTA/2 mM MgCl2/0.2% Triton X-100.…”

Section: Methodsmentioning

confidence: 99%

Identification of ubiquitous high-molecular-mass, heat-stable microtubule-associated proteins (MAPs) that are related to the Drosophila 205-kDa MAP but are not related to the mammalian MAP-4.

Kimble¹,

Khodjakov²,

Kuriyama³

1992

Proc. Natl. Acad. Sci. U.S.A.

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Identification of ubiquitous high-molecular-mass, heat-stable microtubule-associated proteins (MAPs) The role of microtubules in a variety of cellular processes appears to be regulated, in part, by the many proteins that associate with microtubules. Microtubule-associated proteins (MAPs) have been characterized in a number of different systems and represent a diverse group ofproteins that are generally classified according to their apparent molecular masses or biochemical properties (for reviews, see refs. 1-4). The MAP-2, MAP-4, and T classes include proteins that are fibrous in structure and are stable at elevated temperatures. Of these, the largest number of proteins so far described has been in the MAP-4 class, members of which range from 180 to 240 kDa and which includes mouse MAP-4 (5), HeLa 210-kDa MAP (6, 7), Drosophila 205-kDa MAP (8), the 190-kDa MAPs identified in bovine adrenal gland (9) and rat liver (10), and rat MAP-3 (11). Complete or partial DNA sequence has now been reported for the genes encoding four of the MAP-4 class of proteins (12-15). Comparison of the predicted amino acid sequences of the HeLa, bovine, and mouse MAP-4 proteins has shown an overall sequence identity of -70%; the conserved residues of these three proteins are clustered in four regions, one of which includes the microtubule-binding domain (15). In contrast, the predicted amino acid sequence of the Drosophila 205-kDa MAP is not similar to either the mammalian MAP-4 proteins or to any other MAP for which sequence data is available (12).Using a monoclonal antibody (mAb; AX3) raised against microtubule-organizing centers from the cellular slime mold Dictyostelium discoideum, we have identified a highmolecular mass, heat-stable MAP that occurs in species ranging from Dictyostelium to humans (16). We show here that the proteins recognized by mAb AX3 in mouse and human are distinct from the previously identified MAP-2 and MAP-4 proteins. However, the AX3 antigens in Dictyostelium and mammals are immunologically related to the 205-kDa MAP of Drosophila, suggesting that the 205-kDa MAP and the AX3 antigens define a distinct class of heat-stable MAPs. MATERIALS AND METHODSAntibodies. The AX3 mAb was raised against an isolated nucleus/nucleus-associated body (the primary microtubuleorganizing center of Dictyostelium) fraction from the Ax-3 strain of Dictyostelium discoideum. The production and preliminary characterization of this mAb are described elsewhere (16).The antibodies against mouse MAP-4, HeLa 210-kDa MAP, Drosophila 205-kDa MAP, and mouse MAP-2 used in this study were provided by J. B. Olmsted (17), G. G. Borisy (6), L. S. B. Goldstein (8), and L. I. Binder (University of Alabama, Birmingham), respectively. Anti-tubulin antibodies used were either an antibody raised against yeast tyrosinated a-tubulin (from J. V. Kilmartin) (18) or an anti-chicken (-tubulin mAb (Amersham).Cell Culture and Protein Fractionation. Dictyostelium cells grown in suspension were collected by centrifugation and resuspended in 8-10 vol of homogeni...

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Chapter 15 Probing the Mechanisms of Mitosis with Dictyostelium discoideum

Cited by 19 publications

References 59 publications

The Actomyosin Cytoskeleton of Amoebae of the Cellular Slime MoldsAcrasis roseaandProtostelium mycophaga: Structure, Biochemical Properties, and Function

The Actomyosin Cytoskeleton of Amoebae of the Cellular Slime MoldsAcrasis roseaandProtostelium mycophaga: Structure, Biochemical Properties, and Function

Requirements for Hirano Body Formation

Identification of ubiquitous high-molecular-mass, heat-stable microtubule-associated proteins (MAPs) that are related to the Drosophila 205-kDa MAP but are not related to the mammalian MAP-4.

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