Functional Elimination of Calmodulin within the Nucleus by Targeted Expression of an Inhibitor Peptide

Wang, Jiahong; Campos, Begoña; Jamieson, Gordon A.; Kaetzel, Marcia A.; Dedman, John R.

doi:10.1074/jbc.270.51.30245

Cited by 76 publications

(80 citation statements)

References 29 publications

(17 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CaMBP4 is a nuclear protein that contains four repeats of the M13 calmodulin-binding peptide from the skeletal muscle myosin light chain kinase; it binds to and inactivates the nuclear calcium/CaM complex (30) and has been used previously to identify nuclear calciumregulated genes (18,19,23). Expression of CaMBP4, as well as that of hrGFP after infection with rAAV-CaMBP4 and rAAVhrGFP, was readily detectable immunocytochemically in 80 -95% of the viable cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

Nuclear Calcium Signaling Controls Methyl-CpG-binding Protein 2 (MeCP2) Phosphorylation on Serine 421 following Synaptic Activity

Buchthal

Lau

Weiß

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: MeCP2 is required for synaptogenesis and proper development of neuronal circuits. Results: MeCP2 phosphorylation on serine 421 is controlled by nuclear calcium signaling activating nuclear CaMKII. Conclusion: This defines a novel pathway through which nuclear calcium regulates synaptic activity-driven genomic responses. Significance: Nuclear calcium modulates the function of a key regulator of neuronal circuit development.

show abstract

Section: Resultsmentioning

confidence: 99%

Nuclear Calcium Signaling Controls Methyl-CpG-binding Protein 2 (MeCP2) Phosphorylation on Serine 421 following Synaptic Activity

Buchthal

Lau

Weiß

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…A synthetic gene that encodes the calmodulin-binding sequence of myosin-light chain kinase [24,25] (kindly provided by Marcia Kaetzel and John Dedman, University of Cincinnati) was used. The construct contains four tandem calmodulin-binding peptide (CaMBP) repeats.…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

“…A peptide corresponding to the calmodulin-binding domain of myosin-light chain kinase specifically inhibits calmodulin function in cells [24,25]. By selectively targeting this construct to the nucleus or plasma membrane, we are able to inhibit calmodulin function in discrete subcellular domains [25].…”

Section: Effect Of Calmodulin On Iqgap1-stimulated Cell Migrationmentioning

confidence: 99%

Multiple proteins mediate IQGAP1-stimulated cell migration

Mataraza

Jeong

et al. 2007

Cellular Signalling

View full text Add to dashboard Cite

Cell migration, a highly complex physiological phenomenon that requires the co-ordinated and tightly regulated function of several proteins, is mediated by a number of signalling pathways. Elucidation of the molecular mechanisms of cell migration impacts our comprehension of numerous cell functions, ranging from development and immune surveillance to angiogenesis and metastasis. The scaffold protein IQGAP1, which binds multiple proteins and regulates their functions, promotes cell motility. Many of the IQGAP1 binding proteins have been implicated in cell migration. In this study, we employed a multifaceted strategy to identify proteins that contribute to IQGAP1-stimulated cell migration. Using specific IQGAP1 point mutant constructs, an interaction with actin was shown to be essential for IQGAP1 to increase cell migration. In contrast, eliminating the binding of Ca 2+ / calmodulin, but not Ca 2+ -free calmodulin, augmented the ability of IQGAP1 to stimulate cell migration. Consistent with these findings, selective inhibition of calmodulin function at the plasma membrane with a specific peptide inhibitor enhanced cell migration mediated by IQGAP1. Interestingly, immunofluorescence staining and confocal microscopy suggest that localization of Cdc42 at the leading edge is not necessary for maximal migration of epithelial cells. Coupled with the observations that Cdc42 and Rac1 contribute to IQGAP1-stimulated cell migration, these data suggest that IQGAP1 serves as a junction to integrate multiple signalling molecules to facilitate cell migration.

show abstract

“…Plasmid Construction-A synthetic gene that encodes the myosin light chain kinase calmodulin-binding sequence was used (15). The Flag-tagged construct, which comprises four tandem calmodulin-binding peptide (CaMBP) repeats, is termed CaMBP4-Flag.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, we adopted a complementary strategy to inhibit calmodulin. Transient transfection of an inhibitor peptide derived from muscle myosin light-chain kinase into mammalian cells blocks calmodulin function (15). The CaMBP was tagged with Flag and EYFP.…”

Section: Development Of Cambps To Specifically Inhibit Calmodulin In mentioning

confidence: 99%

Calmodulin Regulates the Transcriptional Activity of Estrogen Receptors

Sacks

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

The steroid hormone estrogen elicits biological effects in cells by binding to and activating the estrogen receptor (ER). Estrogen binding induces a conformational change in the receptor, inducing nuclear translocation and transcriptional activation of ER. The ubiquitous Ca 2؉ -binding protein calmodulin has been shown to interact directly with ER and enhance its stability. To further elucidate the functional sequelae of the association between calmodulin and ER, we examined the effect on ER transcriptional activation of specifically inhibiting calmodulin. The cell-permeable calmodulin antagonist CGS9343B prevented estrogen-induced transcriptional activation by ER, without altering basal transcription. The inhibition was dose-dependent and independent of the time of estrogen stimulation. To validate these findings, calmodulin function was also neutralized by targeted expression of a specific inhibitor peptide. By inserting localization signals, the inhibitor peptide was selectively targeted to different subcellular domains. Inactivation of calmodulin function in the nucleus virtually eliminated estrogen-stimulated ER transcriptional activation. By contrast, when membrane calmodulin was specifically neutralized, estrogen-stimulated transcriptional activation by ER was only slightly attenuated. Importantly, the inhibitor peptides did not significantly reduce the amount of ER in the cells. Together, these data demonstrate that calmodulin is a fundamental component of ER transcriptional activation.The classic steroid hormone estrogen promotes the proliferation of both normal and malignant breast epithelial cells and shortens the cell cycle. Estrogen mediates its biological effects in cells through the estrogen receptor (ER), 1 a member of the nuclear receptor family of ligand-dependent transcription factors (reviewed in Refs. 1 and 2). Analogous to other steroid hormone receptors, ER is an intracellular transcription factor composed of six domains. Estrogen binding to the C-terminal hormone-binding domain induces conformational changes in ER, thereby promoting its dimerization and nuclear localization. The DNA-binding domain of the activated ER binds to DNA sequences, termed estrogen response elements, found in the regulatory regions of target genes. Several factors, including coactivators, corepressors, and integrator proteins, are important in ER-mediated transcription (reviewed in Refs. 3 and 4). It is becoming apparent that transcriptional regulation requires the recruitment by ER of multiple, distinct proteins that cooperate to achieve the required response (3). These factors can alter the magnitude of cellular responses to estrogen.There are yet additional factors that modulate ER function. For example, ER interacts with members of the heat-shock protein family (1), and dissociation of heat-shock protein seems to be necessary for ER to activate transcription. One of the major roles of ligand binding is to change the nature of proteinprotein interactions between steroid receptors and other proteins (2). Convers...

show abstract

Functional Elimination of Calmodulin within the Nucleus by Targeted Expression of an Inhibitor Peptide

Cited by 76 publications

References 29 publications

Nuclear Calcium Signaling Controls Methyl-CpG-binding Protein 2 (MeCP2) Phosphorylation on Serine 421 following Synaptic Activity

Nuclear Calcium Signaling Controls Methyl-CpG-binding Protein 2 (MeCP2) Phosphorylation on Serine 421 following Synaptic Activity

Multiple proteins mediate IQGAP1-stimulated cell migration

Calmodulin Regulates the Transcriptional Activity of Estrogen Receptors

Contact Info

Product

Resources

About