The Role of Calmodulin Recruitment in Ca2+ Stimulation of Adenylyl Cyclase Type 8

Simpson, Rachel E.; Ciruela, Antonio; Cooper, Dermot M.F.

doi:10.1074/jbc.m510992200

Cited by 36 publications

(61 citation statements)

References 42 publications

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“…2D). The reduced responsiveness of AC8M1 to CCE has been attributed to the deletion of an N-terminal calmodulin (CaM) binding domain (residues 34-51), which precludes pre-recruitment of CaM (although additional factors are not excluded) (Gu and Cooper, 1999;Simpson et al, 2006) (Fig. 2D).…”

Section: Ac8 Enhances the Cortical Actin Signalmentioning

confidence: 99%

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Ayling

Briddon

Halls

et al. 2012

Journal of Cell Science

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SummaryThe central and pervasive influence of cAMP on cellular functions underscores the value of stringent control of the organization of adenylyl cyclases (ACs) in the plasma membrane. Biochemical data suggest that ACs reside in membrane rafts and could compartmentalize intermediary scaffolding proteins and associated regulatory elements. However, little is known about the organization or regulation of the dynamic behaviour of ACs in a cellular context. The present study examines these issues, using confocal image analysis of various AC8 constructs, combined with fluorescence recovery after photobleaching and fluorescence correlation spectroscopy. These studies reveal that AC8, through its N-terminus, enhances the cortical actin signal at the plasma membrane; an interaction that was confirmed by GST pull-down and immunoprecipitation experiments. AC8 also associates dynamically with lipid rafts; the direct association of AC8 with sterols was confirmed in Förster resonance energy transfer experiments. Disruption of the actin cytoskeleton and lipid rafts indicates that AC8 tracks along the cytoskeleton in a cholesterol-enriched domain, and the cAMP that it produces contributes to sculpting the actin cytoskeleton. Thus, an adenylyl cyclase is shown not just to act as a scaffold, but also to actively orchestrate its own micro-environment, by associating with the cytoskeleton and controlling the association by producing cAMP, to yield a highly organized signalling hub.

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Section: Ac8 Enhances the Cortical Actin Signalmentioning

confidence: 99%

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Ayling

Briddon

Halls

et al. 2012

Journal of Cell Science

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“…Nine transmembrane AC isoforms have been identified, four of which are sensitive to Ca 2ϩ ; the cation stimulates AC1 and AC8 via calmodulin and inhibits AC5 and AC6 directly. AC8 is an archetypal calmodulin-stimulated enzyme, employing a disinhibitory activation mechanism (Gu and Cooper, 1999;Simpson et al, 2006). However, AC8 is extremely discerning in its responsiveness to elevation of the cytoplasmic Ca 2ϩ concentration ([Ca 2ϩ ] i ).…”

Section: ϩmentioning

confidence: 99%

Capacitative Ca²⁺Entry via Orai1 and Stromal Interacting Molecule 1 (STIM1) Regulates Adenylyl Cyclase Type 8

Martin¹,

Willoughby²,

Ciruela³

et al. 2009

Mol Pharmacol

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Capacitative Ca 2ϩ entry (CCE), which occurs through the plasma membrane as a result of Ca 2ϩ store depletion, is mediated by stromal interacting molecule 1 (STIM1), a sensor of intracellular Ca 2ϩ store content, and the pore-forming component Orai1. However, additional factors, such as C-type transient receptor potential (TRPC) channels, may also participate in the CCE apparatus. To explore whether the store-dependent Ca 2ϩ entry reconstituted by coexpression of Orai1 and STIM1 has the functional properties of CCE, we used the Ca 2ϩ -calmodulin stimulated adenylyl cyclase type 8 (AC8), which responds selectively to CCE, whereas other modes of Ca 2ϩ entry, including those activated by arachidonate and the ionophore ionomycin, are ineffective. In addition, the Ca 2ϩ entry mediated by previous CCE candidates, diacylglycerol-activated TRPC channels, does not activate AC8.Here, we expressed Orai1 and STIM1 in HEK293 cells and saw a robust increment in CCE, and a proportional increase in CCEstimulated AC8 activity. Inhibitors of the CCE assembly process ablated the effects on cyclase activity in both AC8-overexpressing HEK293 cells and insulin-secreting MIN6 cells endogenously expressing Ca 2ϩ -sensitive AC isoforms. AC8 is believed to be closely associated with the source of CCE; indeed, not only were AC8, Orai1, and STIM1 colocalized at the plasma membrane but also all three proteins occurred in lipid rafts. Together, our data indicate that Orai1 and STIM1 can be integral components of the cAMP and CCE microdomain associated with adenylyl cyclase type 8.

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“…AC 5, 6, and 8 NT bind Gbg (Crossthwaite et al, 2006;Gao et al, 2007). AC5NT associates with the guanine nucleotide exchange factor Ric8a (Wang et al, 2007), whereas AC8NT binds the phosphatase PP2A (Crossthwaite et al, 2006), as well as facilitating AC8 stimulation by calmodulin (Simpson et al, 2006). AC NT binding sites for protein kinase C also vary by isoform (Lai et al, 1999;Chou et al, 2004;Crossthwaite et al, 2006;Simpson et al, 2006;Wang et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…AC5NT associates with the guanine nucleotide exchange factor Ric8a (Wang et al, 2007), whereas AC8NT binds the phosphatase PP2A (Crossthwaite et al, 2006), as well as facilitating AC8 stimulation by calmodulin (Simpson et al, 2006). AC NT binding sites for protein kinase C also vary by isoform (Lai et al, 1999;Chou et al, 2004;Crossthwaite et al, 2006;Simpson et al, 2006;Wang et al, 2007). The NT of AC 2, 5, 6, and 9 can interact with macromolecular signaling scaffolds such as A-kinase anchoring proteins (AKAPs) that facilitate spatiotemporal control of AC activity (Piggott et al, 2008;Efendiev et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Adenylyl Cyclase 5 Regulation by GβγInvolves Isoform-Specific Use of Multiple Interaction Sites

et al. 2015

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Adenylyl cyclase (AC) converts ATP into cyclic AMP (cAMP), an important second messenger in cell signaling. Heterotrimeric G proteins and other regulators are important for control of AC activity. Depending on the AC isoform, Gbg subunits can either conditionally stimulate or inhibit cAMP synthesis. We previously showed that the Ga s -bg heterotrimer binds to the N terminus (NT) of type 5 AC (AC5). We now show that Gbg binds to the NT of a wide variety of AC isoforms. We hypothesized that Gbg/AC5 interactions involving inactive heterotrimer and Gbg stimulation of AC5 were separable events. Mutations of the Gbg "hotspot" show that this site is necessary for AC5 stimulation but not for interactions with the first 198 aa of AC5NT, which is a G protein scaffolding site. This contrasts with AC6, where the Gbg hotspot is required for both interactions with AC6NT and for stimulation of AC6. Additionally, the SIGK hotspot peptide disrupts Gbg regulation of AC isoforms 1, 2, and 6, but not AC5. Gbg also binds the C1/C2 catalytic domains of AC5 and AC6. Finally, cellular interactions with full-length AC5 depend on multiple sites on Gbg. This suggests an isoform-specific mechanism in which bound Gbg at the AC5NT is ideally situated for spatiotemporal control of AC5. We propose Gbg regulation of AC involves multiple binding events, and the role of the AC NT for mechanisms of regulation by heterotrimeric G protein subunits is isoform-specific.

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The Role of Calmodulin Recruitment in Ca2+ Stimulation of Adenylyl Cyclase Type 8

Abstract: Ca

Cited by 36 publications

References 42 publications

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Capacitative Ca²⁺Entry via Orai1 and Stromal Interacting Molecule 1 (STIM1) Regulates Adenylyl Cyclase Type 8

Adenylyl Cyclase 5 Regulation by GβγInvolves Isoform-Specific Use of Multiple Interaction Sites

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The Role of Calmodulin Recruitment in Ca2+ Stimulation of Adenylyl Cyclase Type 8

Abstract: Ca

Cited by 36 publications

References 42 publications

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Adenylyl cyclase AC8 directly controls its micro-environment by recruiting the actin cytoskeleton in a cholesterol-rich milieu

Capacitative Ca2+Entry via Orai1 and Stromal Interacting Molecule 1 (STIM1) Regulates Adenylyl Cyclase Type 8

Adenylyl Cyclase 5 Regulation by GβγInvolves Isoform-Specific Use of Multiple Interaction Sites

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Capacitative Ca²⁺Entry via Orai1 and Stromal Interacting Molecule 1 (STIM1) Regulates Adenylyl Cyclase Type 8