Mechanical control of cyclic AMP signalling and gene transcription through integrins

Meyer, Christian J.; Alenghat, Francis J.; Rim, Paul; Fong, Jim C.; Fabry, Ben; Ingber, Donald E.

doi:10.1038/35023621

Cited by 236 publications

(215 citation statements)

References 21 publications

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“…Les premiers gènes mécanosensi-bles découverts ont été des gènes de « réponse à un stress mécanique », au sens où ils fonctionnent de manière à adapter la structure du tissu à son environnement mécanique, par exemple en le renforçant par le déclenchement de divisions cellulaires, ou par l'expression de protéines composants le cytosquelette [2,3]. Dans ce cadre, on a mis en évi-dence qu'un certain nombre de facteurs de transcription, dont NF-κB, étaient sensibles aux contraintes mécaniques, dans ce cas leur translocation nucléaire est induite mécaniquement [4].…”

Section: Gènes Mécanosensiblesunclassified

À la recherche des capteurs mécaniques moléculaires de la cellule

Desprat

Farge

2007

Med Sci (Paris)

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Section: Gènes Mécanosensiblesunclassified

À la recherche des capteurs mécaniques moléculaires de la cellule

Desprat

Farge

2007

Med Sci (Paris)

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“…35,39 From a biochemical perspective, local modulation of integrin receptors affects cell behavior because applying mechanical stresses alters the expression of cyclic AMP as well as induces local recruitment of mRNA and ribosomes to focal adhesions. 10,65 Membrane receptor signaling and gene transcription are activated in a force-dependent manner by applying magnetic twisting forces to these receptors via ferromagnetic beads coated with specific receptor ligands (Fig. 4).…”

Section: Mechanotransductionmentioning

confidence: 99%

“…4). 65,112 Magnetically twisting cell surface adhesion receptors that are clustered (oligomerized) within focal adhesion complexes result in a rapid increase in the production of the second messenger, cyclic AMP. This increase in cyclic AMP leads to downstream signaling events resulting in the activation of gene transcription, as confirmed by demonstrating that magnetic activation of a reporter gene that encodes the enzyme, β-galactosidase, is driven by cyclic AMP-response elements.…”

Section: Mechanotransductionmentioning

confidence: 99%

Nanoscale Intracellular Organization and Functional Architecture Mediating Cellular Behavior

LeDuc

Bellin

2006

Ann Biomed Eng

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Abstract-Cells function based on a complex set of interactions that control pathways resulting in ultimate cell fates including proliferation, differentiation, and apoptosis. The interworkings of this immensely dense network of intracellular molecules are influenced by more than random protein and nucleic acid distribution where their interactions culminate in distinct cellular function. By probing the design of these biological systems from an engineering perspective, researchers can gain great insight that will aid in building and utilizing systems that are on this size scale where traditional large-scale rules may fail to apply. The organized interaction and gradient distribution in intracellular space imply a structural architecture that modulates cellular processes by influencing biochemical interactions including transport and binding-reactions. One significant structure that plays a role in this modulation is the cell cytoskeleton. Here, we discuss the cytoskeleton as a central and integrating functional structure in influencing cell processes and we describe technology useful for probing this structure. We explain the nanometer scale science of cytoskeletal structure with respect to intracellular organization, mechanotransduction, cytoskeletal-associated proteins, and motor molecules, as well as nano-and microtechnologies that are applicable for experimental studies of the cytoskeleton. This biological architecture of the cytoskeleton influences molecular, cellular, and physiological processes through structured multimodular and hierarchical principles centered on these functional filaments. Through investigating these organic systems that have evolved over billions of years, understanding in biology, engineering, and nanometer-scaled science will be advanced.

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“…Specifically, magnetic twisting of integrins that are clustered within focal adhesions results in a rapid increase in the production of the second messenger, adenosine 3′,5′-monophosphate (cAMP), inside the cell [15]. This increase in cAMP induces downstream signaling through protein kinase A and leads to the formation of a protein transcription complex that binds to an eight-nucleotide DNA sequence, known as the cAMP regulatory element (CRE), and promotes transcription of neighboring genes [18].…”

Section: Nih Public Accessmentioning

confidence: 99%

“…Previous work from our laboratory has identified that intracellular signaling and gene transcription can be activated in a force-dependent manner by magnetically applying twisting forces to cell surface integrin receptors using ferromagnetic microbeads (1-5 µm diameter) coated with specific receptor ligands, such as synthetic RGD-containing peptides from the cell binding region of the ECM protein fibronectin [4], [11], [15]. Specifically, magnetic twisting of integrins that are clustered within focal adhesions results in a rapid increase in the production of the second messenger, adenosine 3′,5′-monophosphate (cAMP), inside the cell [15].…”

Section: Nih Public Accessmentioning

confidence: 99%

Magnetic Cellular Switches

et al. 2004

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This paper focuses on the development of magnetic cellular switches to enable magnetic control of intracellular functions in living mammalian cells, including receptor signal transduction and gene transcription. Our approach takes advantage of the mechanosensitivity of adenosine 3′,5′-monophosphate (cAMP) induction and downstream transcription controlled by the cAMP regulatory element (CRE) to engineer gene constructs that optically report gene expression in living cells. We activate transcription of these gene reporters by applying magnetic (mechanical) stress to magnetic microbeads bound to cell surface integrin receptors. In these gene reporter constructs, CRE motifs drive the expression of fluorescent proteins or enzymes that produce fluorescent products, such as DsRed and β-lactamase (BLA), respectively. We demonstrate that a chemical inducer of cAMP (forskolin) increases expression of CRE-DsRed in living cells. More importantly, a threefold increase in CRE-BLA expression is induced by application of mechanical stress to magnetic microbeads (4.5 µm) bound to cell surface integrin receptors. Induction of cAMP could be detected within 5 min using a protein fragment complementation assay involving interactions between the KID and KIX domains of the CRE binding protein linked to complementary halves of the BLA enzyme. These studies confirm that application of magnetic stress to integrins induces gene transcription by activating the cAMP-dependent transcription factor CREB. Ongoing studies focus on optimizing sensitivity and reducing signal-to-noise by establishing stable cell lines that express these gene reporters. These studies collectively demonstrate the feasibility of using magnetic technologies to control function in living mammalian cells and, hence, support the possibility of developing magnetically-actuated cellular components for use in future micro-and nanotechnologies.

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Mechanical control of cyclic AMP signalling and gene transcription through integrins

Cited by 236 publications

References 21 publications

À la recherche des capteurs mécaniques moléculaires de la cellule

À la recherche des capteurs mécaniques moléculaires de la cellule

Nanoscale Intracellular Organization and Functional Architecture Mediating Cellular Behavior

Magnetic Cellular Switches

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