Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments.

Hubbard, Bruce D.; Lazarides, Elias

doi:10.1083/jcb.80.1.166

Cited by 198 publications

(107 citation statements)

References 21 publications

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“…In this experimental condition, nucleotide triphosphate, nucleotide diphosphate, nucleotide thiotriphosphate, and Ca 2ϩ produced no detectable changes in the outer PD rings (Table 1). In summary, the 5-nm filament was relatively insoluble, and the solubility properties resemble those of intermediate filaments such as desmin and vimentin (Hubbard and Lazarides, 1979;Lazarides and Granger, 1982), although the dimension is different (intermediate filaments are 10 nm in diameter). In addition, the bundle of filaments was so rigid that it could not be broken by freezing and thawing.…”

Section: The Bundle Of the Filament Is A Relatively Insoluble Structurementioning

confidence: 99%

Novel Filaments 5 nm in Diameter Constitute the Cytosolic Ring of the Plastid Division Apparatus

Miyagishima¹,

Takahara²,

Kuroiwa³

2001

The Plant Cell

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The plastid division apparatus (called the plastid-dividing ring) has been detected in several plant and algal species at the constricted region of plastids by transmission electron microscopy. The apparatus is composed of two or three rings: an outer ring in the cytosol, an inner ring in the stroma, and a middle ring in the intermembrane space. The components of these rings are not clear. FtsZ, which forms the bacterial cytokinetic ring, has been proposed as a component of both the inner and outer rings. Here, we present the ultrastructure of the outer ring at high resolution. To visualize the outer ring by negative staining, we isolated dividing chloroplasts from a synchronized culture of a red alga, Cyanidioschyzon merolae , and lysed them with nonionic detergent Nonidet P-40. Nonidet P-40 extracted primarily stroma, thylakoids, and the inner and middle rings, leaving the envelope and outer ring largely intact. Negative staining revealed that the outer ring consists of a bundle of 5-nm filaments in which globular proteins are spaced 4.8 nm apart. Immunoblotting using an FtsZ-specific antibody failed to show immunoreactivity in the fraction containing the filament. Moreover, the filament structure and properties are unlike those of known cytoskeletal filaments. The bundle of filaments forms a very rigid structure and does not disassemble in 2 M urea. We also identified a dividing phase-specific 56-kD protein of chloroplasts as a candidate component of the ring. Our results suggest that the main architecture of the outer ring did not descend from cyanobacteria during the course of endosymbiosis but was added by the host cell early in plant evolution. INTRODUCTIONEukaryotic cells contain organelles that are duplicated and inherited by daughter cells during cell division. Among these organelles, mitochondria and plastids are unique in that they contain DNA-protein complexes (nucleoids) and machinery sufficient for protein synthesis. It is generally believed that mitochondria and plastids arose from prokaryotic endosymbionts during eukaryotic evolution. The endosymbiotic theory states that the progenitor of mitochondria was an ␣ -proteobacterium and that the plastid ancestor was a cyanobacterium (Gray, 1992). Mitochondria and plastids multiply by the division of preexisting organelles, as do bacteria (Leech et al., 1981;Kuroiwa, 1982Kuroiwa, , 1991Possingham and Lawrence, 1983; Boffey and Lloyd, 1988). Of the characterized mitochondrial and plastid genomes, however, none has a complete set of genes sufficient for self-replication. Mitochondria that cannot synthesize proteins as a result of large genome deletions (petite mutant) (Attardi and Schatz, 1988) and plastids that lack ribosomes (Hashimoto and Possingham, 1989) still can proliferate. Therefore, it is believed that products from the host (nuclear) genomes perform mitochondrial and plastid division. Nuclear regulation of organelle division remains poorly understood, but in plastid division both the plastid-dividing ring (PD ring) (summarized in Kuroiwa et ...

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Section: The Bundle Of the Filament Is A Relatively Insoluble Structurementioning

confidence: 99%

Novel Filaments 5 nm in Diameter Constitute the Cytosolic Ring of the Plastid Division Apparatus

Miyagishima¹,

Takahara²,

Kuroiwa³

2001

The Plant Cell

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“…One-dimensional SDS PAGE was based on the system of Laemmli (22) as modified and described previously (18). Separating gels contained either 12.5% acrylamide or 10% acrylamide as detailed in the figure legends.…”

Section: Pacementioning

confidence: 99%

Avian lens spectrin: subunit composition compared with erythrocyte and brain spectrin.

Nelson¹,

Granger²,

Lazarides³

1983

The Journal of Cell Biology

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Chicken lens spectrin is composed predominantly of equimolar amounts of two polypeptides with solubility properties similar, but not identical, to erythrocyte spectrin. The larger polypeptide, Mr 240,000 (lens a-spectrin), co-migrates with erythrocyte and brain a-spectrin on one-and two-dimensional SDS polyacrylamide gels and cross-reacts with antibodies specific for chicken erythrocyte a-spectrin; the smaller polypeptide, Mr 235,000 (lens 3"-spectrin), co-migrates with brain 3'-spectrin and does not cross-react with either the a-spectrin antibodies or antibodies specific for chicken erythrocyte/3-spectrin. Minor amounts of polypeptides antigenically related to erythrocyte /3-spectrin with a greater electrophoretic mobility than lens 3"-spectrin are also detected in lens. The equimolar ratio of lens a-and 3"-spectrin is invariantly maintained during the extraction of lens plasma membranes under different conditions, or after immunoprecipitation of whole extracts of lens with erythrocyte a-spectrin antibodies. Two-dimensional peptide mapping reveals that whereas a-spectrins from chicken erythrocytes, brain, and lens are highly homologous, the 3"-spectrins, although related, have some cell-type-specific peptides and are substantially different from erythrocyte 1% spectrin. Thus, the expression of cell-type-specific 3'-and /3-spectrins may be the basis for the assembly of a spectrin-plasma membrane complex whose molecular composition is tailored to the functional requirements of the particular cell-type.The cytoskeleton of vertebrate lens cells is composed of microtubules, actin filaments, and vimentin-type intermediate filaments (5,20, 21,33,34,36). Recent studies have indicated that the lens cytoskeleton may play an important role in lens differentiation. Terminal differentiation of lens fiber cells occurs by a gradual process of cellular elongation as cells are displaced laterally from the periphery of the lens towards the center (reviewed in references 1, 4, 17). During this process there is an increase in the ratio of F-actin to Gactin suggesting that cellular elongation is accompanied by an increase in actin filament assembly (34). An active role of actin filaments in cellular elongation is supported also by the results of studies on lens differentiation in vitro that have shown that the disruption of actin filaments with cytochalasin B or D has an inhibitory effect on this process (28). Although the basis of cellular elongation is not fully understood, it has been suggested that it may involve, at least in part, the interaction between actin filaments and the plasma membrane (34,40). In lens fiber cells, actin filaments have been observed frequently in the subcortical region where they appear to be attached to the plasma membrane (1,33,35,36).We have begun to investigate the polypeptide composition of lens cell plasma membranes in order to identify polypeptides involved in this interaction. Initial studies from this laboratory have shown that a polypeptide serologically related to the a-subunit of e...

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“…Desmin is a muscle specific intermediate filament protein found both in smooth muscle (Hubbard and Lazarides, 1979) and localized at the sarcomeric Z-line in striated muscle (Granger and Lazarides, 1979). Desmin has been detected in both mononuclear (Urlakis and Yorde, 1988) and replicating myoblasts (YablonkaReuveni and Nameroff, 1986;Kaufman and Foster, 1988).…”

Section: Introductionmentioning

confidence: 99%

Analysis of the in vivo myogenic status of chick somites by desmin expression in vitro

Borman

Urlakis

Yorde

1994

Developmental Dynamics

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Expression of the muscle specific intermediate filament protein, desmin, is an early marker for chick somitic myogenesis. Somites are transient, paired, mesodermal structures adjacent to the neural tube which are formed very uniformly in a cranial to caudal fashion. The developmental somitic expression of desmin in vivo has been reported previously (Holtzer et al. [1991] "Frontiers in Muscle Research." New York Elsevier Science, pp 187-207; Borman and Yorde U9941 J. Histochem. Cytochem. 42265-272). Here we explore the ability of those somitic cells which are desmin negative in vivo to successfully carry out a myogenic program of development in the absence of the surrounding embryonic microenvironment. Somites which are known to be overtly desmin negative in the embryo were explanted and cultured on collagen gels for 4 days. Immuno-detection of desmin identified a population of somites that could support desmin positive cells in vitro as well as a population of somites that could not. The cranially located somites must remain in the embryo for a greater length of time than the caudally positioned somites prior to each being able to express desmin in vitro. In embryos of many ages there is also a population of somites unable to support desmin expression in vitro. The rate at which this ability to support somitic desmin expression in vitro progresses caudally in the embryo is significantly greater than the rate at which somites form. Notably, the detected expression of desmin in somites in vitro is parallel to the rate at which overt expression of desmin in vivo is detected. The implication for these observations with regard to the regulation of somitic myogenesis is discussed. o

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Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments.

Cited by 198 publications

References 21 publications

Novel Filaments 5 nm in Diameter Constitute the Cytosolic Ring of the Plastid Division Apparatus

Novel Filaments 5 nm in Diameter Constitute the Cytosolic Ring of the Plastid Division Apparatus

Avian lens spectrin: subunit composition compared with erythrocyte and brain spectrin.

Analysis of the in vivo myogenic status of chick somites by desmin expression in vitro

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