Microtubule assembly and conanavalin A capping in lymphocytes: reappraisal using normal and abnormal human peripheral blood cells.

Oliver, Janet M.; Gelfand, Erwin W.; Pearson, Christine B.; Pfeiffer, Janet R.; Dosch, Hans‐Michael

doi:10.1073/pnas.77.6.3499

Cited by 22 publications

(9 citation statements)

References 15 publications

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“…(42) have shown that colchicine is able to induce macrophages to accumulate phagocytic and pinocytic vesicles as well as Con A receptors into a cap structure at one pole of the cell . The primary mode of action of colchicine has been commonly considered to involve the disassembly of microtubules (4,5,22,27,44). However, colchicine has also been found to affect several other cellular activities (e .g., nucleoside transport and membrane components) (25,34,35).…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation of myosin light chain during capping of mouse T-lymphoma cells.

Bourguignon¹,

Nagpal²,

Hsing³

1981

The Journal of Cell Biology

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Colchicine induces the clustering of at least three different T-lymphoma surface antigens (T200, Thy-1, and gp 69/71) into a cap structure in the absence of any external ligand . In addition, colchicine induces the intracellular accumulation of actin and myosin directly beneath the surface cap structure . We have discovered that myosin molecules (both heavy and light chains) are closely associated with the plasma membrane of T-lymphoma cells. Most importantly, we have found that the 20,000-dalton light chain of lymphocyte myosin is both phosphorylated and preferentially accumulated in the plasma membrane of colchicine-induced capped cells. It is proposed that myosin light chain is directly involved in the activation of membrane-associated actomyosin required for the collection of surface proteins into a cap structure (analogous to muscle cell sliding filament contraction).Redistribution of lymphocyte surface receptors can be induced either with or without the addition of external ligands (15,18,23,29,30,37,41,(46)(47)(48) . During the process of receptor movement, surface components first form small clusters (patches), which then aggregate at one pole of the cell into socalled caps .In the case of ligand-dependent capping, the ligand and its receptors appear to be clustered by a cross-linking event at the cell surface that is highly dependent on the valency of the ligand (37). For example, divalent antibodies against surface immunoglobulin (Ig) are required for the induction of Ig capping; monovalent anti-Ig is unable to cause any redistribution of the Ig receptor (30) . In addition, capping induced by the lectin, concanavalin A (Con A), requires the tetrameric form of Con A (at the proper concentration) since the dimer form (e .g ., succinyl Con A) is not active in cap formation (15,18) . Information concerning ligand-independent capping is very limited . Recently, it has been found that hypertonic media, mitogens, trypsin, and cholinergic drugs are each able to induce surface receptors to form cap structures in the absence of any externally added ligand (29,(46)(47)(48). The molecular mechanisms underlying both ligand-dependent and ligand-independent capping are not yet understood .Previously, we have used a double immunofluorescence labeling technique to simultaneously determine the distribution of certain surface receptors and intracellular actin and myosin in human skin fibroblasts grown in monolayers (9) or in murine lymphoid cells grown in suspension cultures (7,10,11) . The THE JOURNAL of CELL BIOLOGY -VOLUME 91 DECEMBER 1981 889-894

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Section: Discussionmentioning

confidence: 99%

Phosphorylation of myosin light chain during capping of mouse T-lymphoma cells.

Bourguignon¹,

Nagpal²,

Hsing³

1981

The Journal of Cell Biology

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“…Such a concept has come mainly from the study of drug effects on the distribution of cell surface proteins. The drugs used include microtubuledepolymerizing agents such as colchicine, nocodazole, and vinblastine (2,4,6) and microfilament-depolymerizing agents such as cytochalasin B (4,8). The distribution of immunoglobulins and concanavalin A receptors on lymphocytes (3,4,6) and fibroblasts (1) after treatment with such drugs has been extensively studied by fluorescence microscopy.…”

mentioning

confidence: 99%

“…It now has been generally accepted that the lateral mobility and distribution of externally exposed cell surface proteins are regulated in large part by submembranous cytoskeletal elements (1)(2)(3)(4)(5)(6), although there is also some evidence that directed lipid flow in membranes is important as well (7). Such a concept has come mainly from the study of drug effects on the distribution of cell surface proteins.…”

mentioning

confidence: 99%

Effect of vinblastine on distribution of murine leukemia virus-derived membrane-associated antigens.

Satake¹,

McMillan²,

Luftig³

1981

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

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The effect of vinblastine on the distribution of murine leukemia virus-derived membrane-associated antigens was examined by using the indirect immunofluorescence of 3.7%formaldehyde-fixed MJD-54 (Moloney murine leukemia virus-infected) cells. On fixed, non-drug-treated cells, p30 antigen was distributed homogeneously and diffusely over the cell membrane.When cells were incubated with 10 pAM vinblastine for 1 hr before fixation, the distribution of p30 antigen was greatly changed, fluorescence now being collected into poles (cap-like formation). In contrast to this distribution pattern for p3) antigen, gp7O antigen was distributed in a micropunctate pattern on the cell surface, with or without vinblastine pretreatment. These observations indicate that the distribution patterns of p30 and gp7O membrane antigens are completely different and that they are differently controlled by cytoplasmic microtubules. In addition, because the p30 membrane antigen visualized in these studies most likely represents viral Pr65P. precursor molecules which are localized directly under and associated with the plasma membrane, these results suggest that, under special conditions of fixation, it is possible to obtain a cap-like phenomenon for cytoplasmic (internal) membraneoriented proteins.It now has been generally accepted that the lateral mobility and distribution of externally exposed cell surface proteins are regulated in large part by submembranous cytoskeletal elements (1-6), although there is also some evidence that directed lipid flow in membranes is important as well (7). Such a concept has come mainly from the study of drug effects on the distribution of cell surface proteins. The drugs used include microtubuledepolymerizing agents such as colchicine, nocodazole, and vinblastine (2, 4, 6) and microfilament-depolymerizing agents such as cytochalasin B (4, 8). The distribution of immunoglobulins and concanavalin A receptors on lymphocytes (3, 4, 6) and fibroblasts (1) after treatment with such drugs has been extensively studied by fluorescence microscopy.In the case ofcells chronically infected with murine leukemia virus there are three known classes ofcell membrane-associated proteins that are coded for by the viral genome. Two are externally exposed; (i) glycosylated gag (group-specific antigen) gene products of Mr 80 x 103 and 95 x 103 (9-11) which have been described earlier as "Gross cell surface antigens" and shown not to be incorporated into virus particles (12); and (ii) the env gene products gp7O and pl5E. gp7O exists both at the site ofvirus buds (13,14) and over the cell surface (9, 13); pl5E is embedded directly in the membrane (15). In addition, there is an internally exposed cell membraneassociated protein. This is the major gag gene product, Pr65P9, which is the precursor to the group-specific viral core proteins (p30, p15, p12, and plo) (10,16). It has been localized, by electron microscopic studies, to a position just under the cell membrane (16) and, because it hasa hydrophobic NH2terminus, p15 (17),...

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“…This proposal is not without precedent . Abnormal cytoskeleton and membrane properties have already been suggested in lymphocytes from patients with two other forms of severe immune dysfunction (6,25) .…”

Section: Discussionmentioning

confidence: 99%

Purine nucleoside phosphorylase is associated with centrioles and basal bodies.

Oliver¹,

Osborne²,

Pfeiffer³

et al. 1981

The Journal of Cell Biology

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We have localized a fraction of the enzyme, purine nucleoside phosphorylase (PNP), to the centrioles and basal bodies of mammalian, avian, and protozoan cells. Two completely independent methods were used, one based on the ultrastructural cytochemistry of the enzyme activity and one based on immunofluorescence microscopy using an antibody raised in rabbit against purified human PNP. PNP catalyzes the reversible conversion of purine nucleosides and inorganic phosphate to the corresponding purine bases and ribose-1-phosphate . Its partial localization to centrioles and basal bodies raises the possibility that purine compounds are involved in centriole replication and/or in the regulation of microtubule assembly in vivo .No centriolar PNP could be detected in primary skin fibroblast from two infants with severe immunodeficiency disease associated with the absence of soluble PNP . This raises the possibility that defects in centriole function may contribute to the impaired division and maturation of T lymphoid precursor in this inherited disorder .Initially, the immunofluorescence analyses were complicated by a residual centriole-binding antibody that persisted in immunoglobulins from immune animals after complete removal of anti-PNP by affinity chromatography . Binding was abolished by exposure of cells to sodium periodate, indicating that this (and possibly other) "spontaneous" anticentriole antibodies in rabbit serum may be directed against carbohydrates .It has become clear from ultrastructural studies that the majority of microtubules in animal cells grow from distinct regions known as microtubule-organizing centers (MTOCs). These structures, which consist of centrioles and their associated satellite substance, basal bodies, and kinetochores, may be the key to understanding the organization and assembly of cytoplasmic, mitotic spindle, and ciliary microtubules . Nevertheless, little is known of their biochemical composition or function (10, 38).We report here that an enzyme, purine nucleoside phosphorylase (PNP ; purine nucleoside ; orthophosphate ribosyltransferase, EC 2 .4 .2.1) is associated with centrioles and their derivatives, the basal bodies of ciliated cells . This enzyme, which catalyzes the reversible reaction of purine nucleosides (guanosine, inosine, and xanthosine, and their deoxyhomologues) with inorganic phosphate to form free bases and (deoxy)ribose-1-phosphate, has been measured previously in soluble extracts of cells ranging from bacteria to man . Its forward reaction (nu-

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Microtubule assembly and conanavalin A capping in lymphocytes: reappraisal using normal and abnormal human peripheral blood cells.

Cited by 22 publications

References 15 publications

Phosphorylation of myosin light chain during capping of mouse T-lymphoma cells.

Phosphorylation of myosin light chain during capping of mouse T-lymphoma cells.

Effect of vinblastine on distribution of murine leukemia virus-derived membrane-associated antigens.

Purine nucleoside phosphorylase is associated with centrioles and basal bodies.

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