Talin Concentrates to the Midbody Region During Mammalian Cell Cytokinesis

Bellissent‐Waydelich, Anne; Vanier, Marie‐Thérèse; Albigès-Rizo, Corinne; Simon‐Assmann, Patricia

doi:10.1177/002215549904701102

Cited by 18 publications

(15 citation statements)

References 52 publications

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“…Talin is ubiquitously expressed and is found in structures ranging from the myotendinous junction (58) and costameres in skeletal muscle (59), the intercalated discs in cardiac muscle (59), dense plaques in smooth muscle (60), the neuromuscular junction (61), the immunological synapse (62), neuronal growth cones, the synapse (63), phagocytic cups (64), and the mid-body in cytokinesis (65). Talin typically colocalizes with vinculin, and it may be significant that in most of these structures the integrin-talin complex is subject to force induced by actomyosin contraction.…”

Section: Discussionmentioning

confidence: 99%

The Activity of the Vinculin Binding Sites in Talin Is Influenced by the Stability of the Helical Bundles That Make Up The Talin Rod

Patel

Gingras

Bobkov

et al. 2006

Journal of Biological Chemistry

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The talin rod contains ϳ11 vinculin binding sites (VBSs), each defined by hydrophobic residues in a series of amphipathic helices that are normally buried within the helical bundles that make up the rod. Consistent with this, talin failed to compete for binding of the vinculin Vd1 domain to an immobilized talin polypeptide containing a constitutively active VBS. However, talin did bind to GST-Vd1 in pull-down assays, and isothermal titration calorimetry measurements indicate a K d of ϳ9 M. Interestingly, Vd1 binding exposed a trypsin cleavage site in the talin rod between residues 898 and 899, indicating that there are one or more active VBSs in the N-terminal part of the talin rod. This region comprises a five helix bundle (residues 482-655) followed by a seven-helix bundle (656 -889) and contains five VBSs (helices 4, 6, 9, 11, and 12). The single VBS within 482-655 is cryptic at room temperature. In contrast, talin 482-889 binds Vd1 with high affinity (K d ϳ 0.14 M), indicating that one or more of the four VBSs within 656 -889 are active, and this likely represents the vinculin binding region in intact talin. In support of this, hemagglutinin-tagged talin 482-889 localized efficiently to focal adhesions, whereas 482-655 did not. Differential scanning calorimetry showed a strong negative correlation between Vd1 binding and helical bundle stability, and a 755-889 mutant with a more stable fold bound Vd1 much less well than wild type. We conclude that the stability of the helical bundles that make up the talin rod is an important factor determining the activity of the individual VBSs.The cytoskeletal protein talin is of a number of proteins including filamin (1), ␣-actinin (2), tensin (3, 4), and ILK (5) implicated in coupling integrins to F-actin in cellular junctions with the extracellular matrix. Talin was originally described as a protein localized in cell extracellular matrix junctions (focal adhesions (FAs)) 2 and membrane ruffles in cultured cells (6) and was subsequently shown to bind integrins in gel filtration studies (7). Its role as a key component of FAs is supported by antibody microinjection experiments (8, 9), laser ablation studies (10), antisense RNA down-regulation (11), and Tln gene disruption studies in cells in culture (12)(13)(14). Knock-out studies in mouse (15), Drosophila (16), and Caenorhabditis elegans (17) confirm that talin is essential for a wide spectrum of integrin-mediated developmental events including gastrulation in the mouse (15), although the situation is complicated by the discovery of a second talin gene (Tln2) in mammals (18). Talin is an elongated (ϳ60 nm) flexible protein (19), reportedly an anti-parallel dimer (20). The globular N-terminal talin head contains a FERM domain (residues 86 -400) with binding sites for the cytoplasmic domains of the integrin ␤-subunit (21, 22) and also layilin (a hyaluronan receptor) (23), F-actin (24), and two signaling proteins, focal adhesion kinase (23) and the type 1␥661 isoform of PI4Ј5Ј-kinase (25-29), both of which are implicated i...

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Section: Discussionmentioning

confidence: 99%

The Activity of the Vinculin Binding Sites in Talin Is Influenced by the Stability of the Helical Bundles That Make Up The Talin Rod

Patel

Gingras

Bobkov

et al. 2006

Journal of Biological Chemistry

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“…It could be that the furrow formation between the dividing cells is more dependent on ␤1 integrin in chondrocytes than in other cell types. The furrow was shown to contain an increased amount of integrin-associated, actin-binding proteins such as ␣-actinin (Fujiwara et al 1978) and talin (Bellissent-Waydelich et al 1999). We could demonstrate that ␤1 integrin subunits localize to the cleavage furrow of dividing chondrocytes, where they might either help to anchor the F-actin ring to the membrane or alternatively induce contraction to complete cell division.…”

Section: ␤1 Integrins Regulate Cytokinesis Of Chondrocytes and Their mentioning

confidence: 99%

β1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis

Aszódi¹,

Hunziker²,

Brakebusch³

et al. 2003

Genes Dev.

292

275

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␤1 integrins are highly expressed on chondrocytes, where they mediate adhesion to cartilage matrix proteins. To assess the functions of ␤1 integrin during skeletogenesis, we inactivated the ␤1 integrin gene in chondrocytes. We show here that these mutant mice develop a chondrodysplasia of various severity. ␤1-deficient chondrocytes had an abnormal shape and failed to arrange into columns in the growth plate. This is caused by a lack of motility, which is in turn caused by a loss of adhesion to collagen type II, reduced binding to and impaired spreading on fibronectin, and an abnormal F-actin organization. In addition, mutant chondrocytes show decreased proliferation caused by a defect in G1/S transition and cytokinesis. The G1/S defect is, at least partially, caused by overexpression of Fgfr3, nuclear translocation of Stat1/Stat5a, and up-regulation of the cell cycle inhibitors p16 and p21. Altogether these findings establish that ␤1-integrin-dependent motility and proliferation of chondrocytes are mandatory events for endochondral bone formation to occur.[Keywords: Integrin; fibronectin; endochondral ossification; growth plate; cytokinesis; FGF] Supplemental material is available at http://www.genesdev.org.

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“…5 These studies show that various phosphorylation events of cytoskeletal 6 and focal adhesion proteins [7][8][9][10][11][12] contribute to the premitotic disassembly of focal adhesions, 13 the deconstruction of the actin cytoskeleton of the interphase, 14 and the construction of a new mitotic cytoskeleton. 15 However, despite this dissociation and deconstruction, the mitotic cell remains attached to the substrate. There is direct contact between the plasma membrane and the substrate.…”

Section: Introductionmentioning

confidence: 99%

Measuring cell adhesion forces during the cell cycle by force spectroscopy

Weder¹,

Vörös²,

Giazzon³

et al. 2009

Biointerphases

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Force spectroscopy has been used to measure the adhesion of Saos-2 cells to a glass surface at different phases of the cell cycle. The cells were synchronized in three phases of the cell cycle: G 1 , S, and G 2 M. Cells in these phases were compared with unsynchronized and native mitotic cells. Individual cells were attached to an atomic force microscope cantilever, brought into brief contact with the glass surface, and then pulled off again. The force-distance curves obtained allowed the work and maximum force of detachment as well as the number, amplitude, and position of discrete unbinding steps to be determined. A statistical analysis of the data showed that the number of binding proteins or protein complexes present at the cell surface and their binding properties remain similar throughout the cell cycle. This, despite the huge changes in cell morphology and adhesion that occur as the cells enter mitosis. These changes are rather associated with the changes in cytoskeletal organization, which can be quantified by force spectroscopy as changes in cell stiffness.

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Talin Concentrates to the Midbody Region During Mammalian Cell Cytokinesis

Cited by 18 publications

References 52 publications

The Activity of the Vinculin Binding Sites in Talin Is Influenced by the Stability of the Helical Bundles That Make Up The Talin Rod

The Activity of the Vinculin Binding Sites in Talin Is Influenced by the Stability of the Helical Bundles That Make Up The Talin Rod

β1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis

Measuring cell adhesion forces during the cell cycle by force spectroscopy

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