Calmodulin and Cytoskeleton

Kakiuchi, Shiro; Sobue, Kenji; Fujita, Masataka; Muramoto, Yoshihiko; Kanda, Keiko; Morimoto, Kouichi

doi:10.1007/978-1-4684-4328-8_12

Cited by 10 publications

(12 citation statements)

References 45 publications

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“…The effects of PMA were reversible. A complete reversal of morphological and cytoskeletal changes was attained within 6 hr after the removal of PMA from the culture medium (Fig. 10).…”

mentioning

confidence: 91%

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Tumor promoter induces reorganization of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells.

Sobue

Fujio

Kanda

1988

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

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We have used immunofluorescence, differential-interference-contrast, and interference-reflection microscopy to examine the translocation of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells induced by phorbol 12-myristate 13-acetate (PMA). The two cytoskeletal proteins were observed to localize in dot structures that corresponded to the cell-substratum contact sites (focal contact) of the cytoplasmic surface of the plasma membrane.The induction of these cytoskeletal changes was specific for tumor promoters. High-resolution microscopy revealed that calspectin was intensely concentrated in ring-like structures surrounding actin dots. It was also located within the areas of actin dots, but to a lesser extent. Trifluoperazine and other phenothiazine derivatives inhibited the formation of those dot structures that appeared after the addition of PMA. Some serine protease inhibitors were also demonstrated to influence cytoskeletal changes by PMA. Our results provide evidence that calspectin is closely associated with actin filaments in dot structures induced by PMA. Possible mechanisms for these cytoskeletal changes produced by PMA are discussed.

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“…The effects of PMA were reversible. A complete reversal of morphological and cytoskeletal changes was attained within 6 hr after the removal of PMA from the culture medium (Fig. 10).…”

mentioning

confidence: 91%

“…In fact, biochemical and immunocytochemical studies support the idea that it mainly locates at the inner surface of the plasma membrane (5)(6)(7)(8)(9). In cultured cells, it remains on the cytoskeletal meshwork even after cells are permeabilized with detergent (10,11).…”

mentioning

confidence: 92%

Tumor promoter induces reorganization of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells.

Sobue

Fujio

Kanda

1988

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

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“…Rotary-shadowed images of the interaction demonstrate that brain spectrin (like erythrocyte spectrin tetramer) is bivalent, binding laterally to F-actin in an end-on orientation, thereby cross-linkmg F-actin strands [Glenney et al, 1982bl. Other characteristics shared by brain and erythrocyte spectrin include a binding site for syndein (or ankyrin) [Bennett et al, 1982;Burridge et al, 19821 and a 240,000 M, a-subunit which can bind calmodulin [Kakiuchi et al, 1982;Glenney et al, 1982c;Palfrey et al, 1982;Carlin et al, 19821. In this report we will present a detailed structural and immunological comparison of membrane-associated brain spectrin and erythrocyte spectrin from mouse.…”

Section: Introductionmentioning

confidence: 98%

A spectrin‐like protein from mouse brain membranes: Immunological and structural correlations with erythrocyte spectrin

et al. 1983

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Membrane-associated mouse brain spectrin is a 972,000 M,, 10.5S, (a& tetramer containing two -240,000 M, subunits and two -235,000 M, subunits. Twodimensional [ 1251]trypti~ peptide mapping indicates that these subunits share only limited and equivalent overlap with the a-and 0-subunits of red blood ccll (RBC) spectrin. Both the 220,000 M, 0-subunit of RBC spectrin and the 235,000 M, 0-subunit of brain spectrin are phosphorylated in the intact mouse. In vitro analysis suggests that both are phosphorylated by a CAMP-independent protein kinase. Antibodies against pure native mouse red blood cell spectrin cross-react with brain spectrin, and antibodies against pure brain spectrin cross-react with both the aand 0-subunits of mouse RBC spectrin. Both antibodies have been utilized to localize brain spectrin within distinct cellular entities of the mouse cerebellum. Granule cell neurons of the internal granule layer and Purkinje cell neurons demonstrated intense fluorescence of the cortical cytoplasm immediately adjacent to the plasma membrane and unstained nuclei, when either RBC or brain spectrin antibodies were utilized for staining. The molecular layer of the cerebellum stained only lightly, and oligodendrocytes and astrocytes appeared to have little fluorescence. Therefore, while brain is a tissue rich in nonerythroid spectrin, the concentration of these immunoreactive analogues is quite variable within distinct cellular entities of the cerebellum.

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“…Calmodulin-Sepharose 4B was prepared by the method of Klee and Krinks (15). Calspectin was prepared from bovine brain by a modification (16) of the method of Kakiuchi et al (17). G-actin was prepared from chicken gizzard smooth muscle as described in ref.…”

mentioning

confidence: 99%

“…4 represent the concentration of calspectin in the cytosol fractions not the total amount in the tissues. Our previous study showed that about 30% of calspectin in brain was distributed in the cytosol fraction; the rest was associated with membranes (17). ' It was of interest to check the possible identity of band 1 with microtubule-associated protein (MAP)1 or MAP2 because MAP2 was reported to be a calmodulin-binding (23) for 60 min at 105,000 x g to precipitate F-actin and proteins bound to F-actin.…”

mentioning

confidence: 99%

Calmodulin-binding proteins that interact with actin filaments in a Ca2+-dependent flip-flop manner: survey in brain and secretory tissues.

Sobue¹,

Kanda²,

Adachi³

et al. 1983

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

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Regulatory actions of calmodulin on the contractile apparatus and cytoskeleton of smooth muscle and nonmuscle tissue are mediated by a number of specific calmodulin-binding proteins that bind to F-actin in a flip-flop manner--i.e., they bind to calmodulin or F-actin depending on the presence or absence, respectively, of Ca2+. A survey for such proteins in brain, adrenal gland, and pituitary gland identified six polypeptides on polyacrylamide gels--Mr 340,000 (band 1), Mr 240,000/235,000 doublet (band 2), Mr 150,000 (band 3), Mr 129,000 (band 4), Mr 105,000 (band 5), and Mr 94,000 (band 6)--as flip-flop-regulated calmodulin- and F-actin-binding polypeptides. In addition to these polypeptides, a Mr 58,000 non-flip-flop calmodulin-binding actin-binding polypeptide (band 7) was found in all tissues examined. Band 2 was identified as calspectin (spectrin-related protein; fodrin). The flip-flop regulation of calspectin required the presence of a heat-labile nondialyzable factor contained in a supernatant fraction of brain homogenates. Band 1 was distinct from microtubule-associated proteins (MAPs) 1 and 2. However, when band 1 polypeptide was kept on ice 3 days, it converted to a lower molecular weight doublet that migrated with MAP2 on NaDodSO4 gel electrophoresis. Bands 1 and 2 were found in all tissues examined.

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Calmodulin and Cytoskeleton

Cited by 10 publications

References 45 publications

Tumor promoter induces reorganization of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells.

Tumor promoter induces reorganization of actin filaments and calspectin (fodrin or nonerythroid spectrin) in 3T3 cells.

A spectrin‐like protein from mouse brain membranes: Immunological and structural correlations with erythrocyte spectrin

Calmodulin-binding proteins that interact with actin filaments in a Ca2+-dependent flip-flop manner: survey in brain and secretory tissues.

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