Association of Calmodulin with Cytoskeletal Structures at Different Stages of HeLa Cell Division, Visualized by a Calmodulin–EGFP Fusion Protein

Erent, Muriel; Pagakis, Stamatis N.; Browne, J.Peter; Bayley, Peter M.

doi:10.1006/mcbr.1999.0137

Cited by 14 publications

(12 citation statements)

References 30 publications

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“…There are a number of observations which show the presence of CaM at the intercellular bridge during the process of cell division. Studies in HeLa cells show that calmodulin is associated with the central spindle and regulates the initiation of cytokinesis (Erent et al ., ; Yu et al ., ).…”

Section: Discussionmentioning

confidence: 98%

The calcium binding protein EhCaBP6 is a microtubular-end binding protein inEntamoeba histolytica

et al. 2013

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SummaryThe genome of Entamoeba histolytica encodes several calcium binding proteins and those characterized thus far have been shown to participate predominantly in phagocytosis and endocytosis. Our study showed that EhCaBP6 has two EF-hand domains EFI and EFIII; it can bind Ca 2+ in vitro and undergoes conformational transition on binding Ca 2+ suggesting that it can serve as a calcium signal sensor. EhCaBP6 is localized in the nucleus, cytoplasm and plasma membrane and is sensitive to heat stress. Unlike other Ca 2+ binding proteins that have been studied in E. histolytica, EhCaBP6 is found at microtubule ends and at the intercellular bridge with the microtubules during cytokinesis. Furthermore, increased expression of EhCaBP6 was correlated with a significant increase in the number of microtubular structures suggesting that this protein may regulate chromosome segregation and cytokinesis in E. histolytica.

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Section: Discussionmentioning

confidence: 98%

The calcium binding protein EhCaBP6 is a microtubular-end binding protein inEntamoeba histolytica

et al. 2013

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“…Moreover, direct evidence suggests that calmodulin and MLCK may be involved in activating the contractile ring. Both calmodulin and some isoforms of MLCK are localized to the cleavage furrow in HeLa cells [4–7]. Loss of calmodulin or application of calmodulin inhibitors disrupts or prevents cytokinesis [7,8] and a MLCK inhibitor causes cytokinesis defects in LLCPK1 cells and crane‐fly spermatocytes [9,10].…”

Section: Introductionmentioning

confidence: 99%

Cytokinesis is not controlled by calmodulin or myosin light chain kinase in the Caenorhabditis elegans early embryo

et al. 2007

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Furrow ingression in animal cell cytokinesis is controlled by phosphorylation of myosin II regulatory light chain (mRLC). In Caenorhabditis elegans embryos, Rho-dependent Kinase (RhoK) is involved in, but not absolutely required for, this phosphorylation. The calmodulin effector myosin light chain kinase (MLCK) can also phosphorylate mRLC and is widely regarded as a candidate for redundant function with RhoK. However, our results show that RNA mediated interference against C. elegans calmodulin and candidate MLCKs had no effect on cytokinesis in wild-type or RhoK mutant embryos, ruling out the calmodulin/MLCK pathway as the missing regulator of cytokinesis in the C. elegans early embryo.

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“…Plasmids pN3-XeCaM-EGFP: wild-type Xenopus calmodulin fusion protein with EGFP (CaM-EGFP) mammalian expression vector was prepared as previously described (Erent et al, 1999). pET24d-XeCaM-EGFP: the coding region for XeCaM-EGFP was excised from a pTRE vector previously prepared (Erent et al, 1999) using a partial NcoI-NotI digest. A modified pET24d (cf.…”

Section: Methodsmentioning

confidence: 99%

“…CaM-EGFP was expressed at <5% endogenous CaM level, which was determined in antiCaM antibody Western blots of cell lysates (cf. Erent et al, 1999).…”

Section: Cell Culture (Mdck and Hela Tet-on Cells)mentioning

confidence: 99%

“…4A for a MDCK control), CaM is distributed at the spindle poles in a dense structure around centrosomes and along the pole to kinetochore microtubules without accumulation at the kinetochores. The centrosomal matrix is a complex 3D structure that appears ring-like in projection (Moudjou et al, 1996;Dictenberg et al, 1998;Erent et al, 1999). Fig.…”

Section: Localisation Of Cam-egfp Under Conditions Of Microtubule Perturbationmentioning

confidence: 99%

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Calmodulin-containing substructures of the centrosomal matrix released by microtubule perturbation

Moisoi¹,

Erent

Whyte³

et al. 2002

Journal of Cell Science

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Calmodulin redistribution in MDCK and HeLa cells subjected to microtubule perturbations by antimitotic drugs was followed using a calmodulin-EGFP fusion protein that preserves the Ca2+ affinity, target binding and activation properties of native calmodulin. CaM-EGFP targeting to spindle structures in normal cell division and upon spindle microtubule disruption allows evaluation of the dynamic redistribution of calmodulin in cell division. Under progressive treatment of stably transfected mammalian cells with nocodazole or vinblastine, the centrosomal matrix at the mitotic poles subdivides into numerous small `star-like' structures, with the calmodulin concentrated centrally, and partially distinct from the reduced microtubule mass to which kinetochores and chromosomes are attached. Prolonged vinblastine treatment causes the release of localised calmodulin into a uniform cytoplasmic distribution, and tubulin paracrystal formation. By contrast,paclitaxel treatment of metaphase cells apparently causes limited disassembly of the pericentriolar material into a number of multipolar `ring-like'structures containing calmodulin, each one having multiple attached microtubules terminating in the partially disordered kinetochore/chromosome complex. Thus drugs with opposite effects in either destabilising or stabilising mitotic microtubules cause subdivision of the centrosomal matrix into two distinctive calmodulin-containing structures, namely small punctate`stars' or larger polar `rings' respectively. The `star-like' structures may represent an integral subcomponent for the attachment of kinetochore microtubules to the metaphase centrosome complex. The results imply that microtubules have a role in stabilising the structure of the pericentriolar matrix, involving interaction, either direct or indirect, with one or more proteins that are targets for binding of calmodulin. Possible candidates include the pericentriolar matrix-associated coiled-coil proteins containing calmodulin-binding motifs, such as myosin V, kendrin (PCNT2) and AKAP450.

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Association of Calmodulin with Cytoskeletal Structures at Different Stages of HeLa Cell Division, Visualized by a Calmodulin–EGFP Fusion Protein

Cited by 14 publications

References 30 publications

The calcium binding protein EhCaBP6 is a microtubular-end binding protein inEntamoeba histolytica

The calcium binding protein EhCaBP6 is a microtubular-end binding protein inEntamoeba histolytica

Cytokinesis is not controlled by calmodulin or myosin light chain kinase in the Caenorhabditis elegans early embryo

Calmodulin-containing substructures of the centrosomal matrix released by microtubule perturbation

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