BBA - Molecular Basis of Disease (v.1842, #12PB)

The role of soluble adenylyl cyclase in health and disease by Jochen Buck; Lonny R. Levin (2533-2534).

The second messenger cyclic adenosine 3′,5′-monophosphate (cAMP) regulates a wide range of physiological processes in almost all organisms. cAMP synthesis is catalyzed by adenylyl cyclases (ACs). All ten mammalian AC isoenzymes (AC1–10) belong to AC Class III, which is defined by sequence homologies in the catalytic domains. Nevertheless, the mammalian AC can be separated into two distinct types, nine transmembrane enzymes (tmAC; AC1–9) and one soluble AC (sAC; AC10). tmACs are mainly regulated by heterotrimeric G-proteins as part of the G-protein coupled receptor pathways, while sAC is directly activated by bicarbonate and Ca2 + and acts as a sensor for ATP, Ca2 +, and bicarbonate/CO2/pH at various intracellular locations. Mammalian sAC has been implicated in processes such as sperm activation, glucose metabolism, and prostate and skin cancer, making it a potential therapeutic target, and first sAC-specific inhibitors have been developed. Mammalian sAC appears evolutionarily closer related to microbial Class III ACs than to tmACs, and sAC-like bicarbonate activated ACs are indeed found in lower organisms and can contribute, e.g., to virulence regulation in microbial pathogens. Here, we review work on the architecture, catalysis, and physiological and pharmacological regulation of sAC-like enzymes, with a main focus on the mammalian enzyme. We further compare the biochemical, regulatory, and structural characteristics of sAC-like enzymes to the evolutionarily and structurally related mammalian tmACs, pointing out common features as well as sAC-specific properties and modulators. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Bicarbonate; cAMP; Calcium; Signaling; Metabolic sensor;

As signaling pathways evolve, selection for new functions guides the co-option of existing material. Major transitions in the history of life, including the evolution of eukaryotes and multicellularity, exemplify this process. These transitions provided both strong selection and a plenitude of available material for the evolution of signaling pathways. Mechanisms that evolved to mediate conflict during the evolution of eukaryotes may subsequently have been co-opted during the many independent derivations of multicellularity. The soluble adenylyl cyclase (sAC) signaling pathway illustrates this hypothesis. Class III adenylyl cyclases, which include sAC, are found in bacteria, including the α-proteobacteria. These adenylyl cyclases are the only ones present in eukaryotes but appear to be absent in archaeans. This pattern suggests that the mitochondrial endosymbiosis brought sAC signaling to eukaryotes as part of an intact module. After transfer to the proto-nuclear genome, this module was then co-opted into numerous new functions. In the evolution of eukaryotes, sAC signaling may have mediated conflicts by maintaining metabolic homeostasis. In the evolution of multicellularity, in different lineages sAC may have been co-opted into parallel tasks originally related to conflict mediation. Elucidating the history of the sAC pathway may be relatively straightforward because it is ubiquitous and linked to near universal metabolic by-products (CO2/HCO3 ). Other signaling pathways (e.g., those involving STAT and VEGF) present a greater challenge but may suggest a complementary pattern. The impact of the mitochondrial endosymbiosis on cell signaling may thus have been profound. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Adenylyl cyclases; Evolution; History of life; Phylogeny; STAT; VEGF;

Role of soluble adenylyl cyclase in mitochondria by Federica Valsecchi; Csaba Konrad; Giovanni Manfredi (2555-2560).
The soluble adenylyl cyclase (sAC) catalyzes the conversion of ATP into cyclic AMP (cAMP). Recent studies have shed new light on the role of sAC localized in mitochondria and its product cAMP, which drives mitochondrial protein phosphorylation and regulation of the oxidative phosphorylation system and other metabolic enzymes, presumably through the activation of intra-mitochondrial PKA. In this review article, we summarize recent findings on mitochondrial sAC activation by bicarbonate (HCO3 ) and calcium (Ca2+) and the effects on mitochondrial metabolism. We also discuss putative mechanisms whereby sAC-mediated mitochondrial protein phosphorylation regulates mitochondrial metabolism. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: sAC; Mitochondria; cAMP; PKA; Protein phosphorylation;

The role of soluble adenylyl cyclase in neurite outgrowth by Travis L. Stiles; Michael S. Kapiloff; Jeffrey L. Goldberg (2561-2568).
Axon regeneration in the mature central nervous system is limited by extrinsic inhibitory signals and a postnatal decline in neurons' intrinsic growth capacity. Neuronal levels of the second messenger cAMP are important in regulating both intrinsic growth capacity and neurons' responses to extrinsic factors. Approaches which increase intracellular cAMP in neurons enhance neurite outgrowth and facilitate regeneration after injury. Thus, understanding the factors which affect cAMP in neurons is of potential therapeutic importance. Recently, soluble adenylyl cyclase (sAC, ADCY10), the ubiquitous, non-transmembrane adenylyl cyclase, was found to play a key role in neuronal survival and axon growth. sAC is activated by bicarbonate and cations and may translate physiologic signals from metabolism and electrical activity into a neuron's decision to survive or regenerate. Here we critically review the literature surrounding sAC and cAMP signaling in neurons to further elucidate the potential role of sAC signaling in neurite outgrowth and regeneration. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Axon growth; Regeneration; Retinal ganglion cell; Optic nerve;

Hypercapnic acidosis activates Ca2+ channels and increases intracellular Ca2+ levels in neurons of the locus coeruleus, a known chemosensitive region involved in respiratory control. We have also shown that large conductance Ca2+-activated K+ channels, in conjunction with this pathway, limits the hypercapnic-induced increase in firing rate in locus coeruleus neurons. Here, we present evidence that the Ca2+ current is activated by a HCO3 -sensitive pathway. The increase in HCO3 associated with hypercapnia activates HCO3 -sensitive adenylyl cyclase (soluble adenylyl cyclase). This results in an increase in cyclic adenosine monophosphate levels and activation of Ca2+ channels via cyclic adenosine monophosphate-activated protein kinase A. We also show the presence of soluble adenylyl cyclase in the cytoplasm of locus coeruleus neurons, and that the cyclic adenosine monophosphate analogue db-cyclic adenosine monophosphate increases Ca2+ i. Disrupting this pathway by decreasing HCO3 levels during acidification or inhibiting either soluble adenylyl cyclase or protein kinase A, but not transmembrane adenylyl cyclase, can increase the magnitude of the firing rate response to hypercapnia in locus coeruleus neurons from older neonates to the same extent as inhibition of K+ channels. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Central control of breathing; L-type Ca2+ channel; Panic disorder; KH7; Development; Transmembrane adenylyl cyclase;

Soluble adenylyl cyclase in the eye by Yong S. Lee; Lihua Y. Marmorstein; Alan D. Marmorstein (2579-2583).
Adenylyl cyclases (ACs) are a family of enzymes which convert ATP to cAMP, an essential intermediate in many signal transduction pathways. Of the 10 AC genes in man, 9 fall into the category of transmembrane ACs (tmACs), which associate with G-protein coupled receptors (GPCRs) and are activated by forskolin. The 10th AC, termed soluble AC (sAC) is neither activated by forskolin nor does it interact with GPCRs. Rather, sAC can be found in many compartments within the cell and is activated by bicarbonate. As such, sAC is considered a major sensor of bicarbonate in many tissues. The pathways involving sAC vary in different tissues and organ systems, and are as diverse as facilitating sperm capacitation and regulating pressure in the eye. The role of sAC in the eye has only recently begun to receive significant attention. Here we summarize what is known about the roles of sAC in the eye. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Adenylyl cyclase; Eye; Retina; Glaucoma; Aqueous humor;

Soluble adenylyl cyclase in health and disease by Andreas Schmid; Dimirela Meili; Matthias Salathe (2584-2592).
The second messenger cAMP is integral for many physiological processes. Soluble adenylyl cyclase (sAC) was recently identified as a widely expressed intracellular source of cAMP in mammalian cells. sAC is evolutionary, structurally, and biochemically distinct from the G-protein-responsive transmembranous adenylyl cyclases (tmAC). The structure of the catalytic unit of sAC is similar to tmAC, but sAC does not contain transmembranous domains, allowing localizations independent of the membranous compartment. sAC activity is stimulated by HCO3 , Ca2 + and is sensitive to physiologically relevant ATP fluctuations. sAC functions as a physiological sensor for carbon dioxide and bicarbonate, and therefore indirectly for pH. Here we review the physiological role of sAC in different human tissues with a major focus on the lung. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease, guest edited by J. Buck and L.R. Levin.
Keywords: cAMP; Soluble adenylyl cyclase; sAC; Bicarbonate; Lung; Airways;

Recently published findings indicate that a knockout (KO) of soluble adenylyl cyclase (sAC, also known as AC-10) gene expression in mice leads to defective glucoregulation that is characterized by reduced pancreatic insulin secretion and reduced intraperitoneal glucose tolerance. Summarized here are current concepts regarding the molecular basis for this phenotype, with special emphasis on the potential role of sAC as a determinant of glucose-stimulated insulin secretion. Highlighted is new evidence that in pancreatic beta cells, oxidative glucose metabolism stimulates mitochondrial CO2 production that in turn generates bicarbonate ion (HCO3 ). Since HCO3 binds to and directly stimulates the activity of sAC, we propose that glucose-stimulated cAMP production in beta cells is mediated not simply by transmembrane adenylyl cyclases (TMACs), but also by sAC. Based on evidence that sAC is expressed in mitochondria, there exists the possibility that beta-cell glucose metabolism is linked to mitochondrial cAMP production with consequent facilitation of oxidative phosphorylation. Since sAC is also expressed in the cytoplasm, sAC catalyzed cAMP production may activate cAMP sensors such as PKA and Epac2 to control ion channel function, intracellular Ca2 + handling, and Ca2 +-dependent exocytosis. Thus, we propose that the existence of sAC in beta cells provides a new and unexpected explanation for previously reported actions of glucose metabolism to stimulate cAMP production. It seems possible that alterations of sAC activity might be of importance when evaluating new strategies for the treatment of type 2 diabetes (T2DM), or when evaluating why glucose metabolism fails to stimulate insulin secretion in patients diagnosed with T2DM. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Soluble adenylyl cyclase; Bicarbonate ion; cAMP; Glucose; Insulin secretion;

The steroid hormone aldosterone regulates the reabsorption of water and ions in the kidney and plays a central role in blood pressure regulation and homeostasis. In recent years, the vascular endothelium has been established as an important aldosterone target organ with major implications in renal and cardiovascular health and disease. Different lines of evidence suggest that the calcium- and bicarbonate-activated soluble adenylyl cyclase (sAC) is a novel mediator of aldosterone signaling in both the kidney and vascular endothelium. This review summarizes our current understanding of the molecular mechanisms of sAC gene expression regulation in the kidney and vascular endothelium and outlines the potential clinical implications of sAC in chronic kidney disease and cardiovascular disease. This review is part of a special issue entitled: The role of soluble adenylyl cyclase in health and disease. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Endothelial stiffness; Bicarbonate; cAMP; Gene expression; Spironolactone;

Central role of soluble adenylyl cyclase and cAMP in sperm physiology by Mariano G. Buffone; Eva V. Wertheimer; Pablo E. Visconti; Dario Krapf (2610-2620).
Cyclic adenosine 3′,5′-monophosphate (cAMP), the first second messenger to be described, plays a central role in cell signaling in a wide variety of cell types. Over the last decades, a wide body of literature addressed the different roles of cAMP in cell physiology, mainly in response to neurotransmitters and hormones. cAMP is synthesized by a wide variety of adenylyl cyclases that can generally be grouped in two types: transmembrane adenylyl cyclase and soluble adenylyl cyclases. In particular, several aspects of sperm physiology are regulated by cAMP produced by a single atypical adenylyl cyclase (Adcy10, aka sAC, SACY). The signature that identifies sAC among other ACs, is their direct stimulation by bicarbonate. The essential nature of cAMP in sperm function has been demonstrated using gain of function as well as loss of function approaches. This review unifies state of the art knowledge of the role of cAMP and those enzymes involved in cAMP signaling pathways required for the acquisition of fertilizing capacity of mammalian sperm. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: cAMP; Sperm capacitation; Soluble adenylyl cyclase; Transmembrane adenylyl cyclase;

Soluble adenylyl cyclase of sea urchin spermatozoa by Victor D. Vacquier; Arlet Loza-Huerta; Juan García-Rincón; Alberto Darszon; Carmen Beltrán (2621-2628).
Fertilization, a key step in sexual reproduction, requires orchestrated changes in cAMP concentrations. It is notable that spermatozoa (sperm) are among the cell types with extremely high adenylyl cyclase (AC) activity. As production and consumption of this second messenger need to be locally regulated, the discovery of soluble AC (sAC) has broadened our understanding of how such cells deal with these requirements. In addition, because sAC is directly regulated by HCO3 it is able to translate CO2/HCO3 /pH changes into cAMP levels. Fundamental sperm functions such as maturation, motility regulation and the acrosome reaction are influenced by cAMP; this is especially true for sperm of the sea urchin (SU), an organism that has been a model in the study of fertilization for more than 130 years. Here we summarize the discovery and properties of SU sperm sAC, and discuss its involvement in sperm physiology. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Adenylyl cyclase; Sperm chemotaxis; Micro-domain; Flagellar motility; Sea urchin sperm; Fertilization;

Soluble adenylyl cyclase (sAC) is poised to play multiple physiological roles as an acid/base (A/B) sensor in aquatic organisms. Many of these roles are probably similar to those in mammals; a striking example is the evolutionary conservation of a mechanism involving sAC, carbonic anhydrase and vacuolar H+-ATPase that acts as a sensor system and regulator of extracellular A/B in shark gills and mammalian epididymis and kidney. Additionally, the aquatic environment presents unique A/B and physiological challenges; therefore, sACs from aquatic organisms have likely evolved distinct kinetic properties as well as distinct physiological roles. sACs from aquatic organisms offer an excellent opportunity for studying the evolution of A/B sensing at both the molecular and whole organism levels. Moreover, this information could help understand and predict organismal responses to environmental stress based on mechanistic models.This article is part of a Special Issue entitled “The Role of Soluble Adenylyl Cyclase in Health and Disease,” guest edited by J. Buck and L. R. Levin.
Keywords: Cyclic AMP; Bicarbonate; Acidosis; Alkalosis; Cell signaling; Carbon Dioxide;

Investigation of cAMP microdomains as a path to novel cancer diagnostics by Garrett Desman; Caren Waintraub; Jonathan H. Zippin (2636-2645).
Understanding of cAMP signaling has greatly improved over the past decade. The advent of live cell imaging techniques and more specific pharmacologic modulators has led to an improved understanding of the intricacies by which cAMP is able to modulate such a wide variety of cellular pathways. It is now appreciated that cAMP is able to activate multiple effector proteins at distinct areas in the cell leading to the activation of very different downstream targets. The investigation of signaling proteins in cancer is a common route to the development of diagnostic tools, prognostic tools, and/or therapeutic targets, and in this review we highlight how investigation of cAMP signaling microdomains driven by the soluble adenylyl cyclase in different cancers has led to the development of a novel cancer biomarker. Antibodies directed against the soluble adenylyl cyclase (sAC) are highly specific markers for melanoma especially for lentigo maligna melanoma and are being described as “second generation” cancer diagnostics, which are diagnostics that determine the ‘state’ of a cell and not just identify the cell type. Due to the wide presence of cAMP signaling pathways in cancer, we predict that further investigation of both sAC and other cAMP microdomains will lead to additional cancer biomarkers. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Soluble adenylyl cyclase; cAMP; Microdomain; Cancer; Diagnostics;

Role of soluble adenylyl cyclase in cell death and growth by Yury Ladilov; Avinash Appukuttan (2646-2655).
cAMP signaling is an evolutionarily conserved intracellular communication system controlling numerous cellular functions. Until recently, transmembrane adenylyl cyclase (tmAC) was considered the major source for cAMP in the cell, and the role of cAMP signaling was therefore attributed exclusively to the activity of this family of enzymes. However, increasing evidence demonstrates the role of an alternative, intracellular source of cAMP produced by type 10 soluble adenylyl cyclase (sAC). In contrast to tmAC, sAC produces cAMP in various intracellular microdomains close to specific cAMP targets, e.g., in nucleus and mitochondria. Ongoing research demonstrates involvement of sAC in diverse physiological and pathological processes. The present review is focused on the role of cAMP signaling, particularly that of sAC, in cell death and growth. Although the contributions of sAC to the regulation of these cellular functions have only recently been discovered, current data suggest that sAC plays key roles in mitochondrial bioenergetics and the mitochondrial apoptosis pathway, as well as cell proliferation and development. Furthermore, recent reports suggest the importance of sAC in several pathologies associated with apoptosis as well as in oncogenesis. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Soluble adenylyl cyclase; Apoptosis; Proliferation; Hypertrophy; Mitochondria; cAMP;

Inhibition of soluble adenylyl cyclase increases the radiosensitivity of prostate cancer cells by Avinash Appukuttan; Jan-Paul Flacke; Hanna Flacke; Alexandra Posadowsky; H. Peter Reusch; Yury Ladilov (2656-2663).
Pharmacological modulation of tumor radiosensitivity is a promising strategy for enhancing the outcome of radiotherapy. cAMP signaling plays an essential role in modulating the proliferation and apoptosis of different cell types, including cancer cells. Until now, the regulation of this pathway was restricted to the transmembrane class of adenylyl cyclases. In the present study, the role of an alternative source of cAMP, the intracellular localized soluble adenylyl cyclase (sAC), in the radiosensitivity of prostate cancer cells was investigated. Pharmacological inhibition of sAC activity led to marked suppression of proliferation, lactate dehydrogenase release, and induction of apoptosis. The combination of ionizing radiation with partial suppression of sAC activity (~ 50%) immediately after irradiation synergistically inhibited proliferation and induced apoptosis. Overexpression of sAC in normal prostate epithelial PNT2 cells increased the cAMP content and accelerated cell proliferation under control conditions. The effects of radiation were significantly reduced in transformed PNT2 cells compared with control cells. Analysis of the underlying cellular mechanisms of sAC-induced radioresistance revealed the sAC-dependent activation of B-Raf/ERK1/2 signaling. In agreement with this finding, inhibition of ERK1/2 in prostate cancer cells enhanced the cytotoxic effect of irradiation. In conclusion, the present study suggests that sAC-dependent signaling plays an important role in the radioresistance of prostate cancer cells. This article is part of a Special Issue entitled: The role of soluble adenylyl cyclase in health and disease.
Keywords: Prostate cancer; Radioresistance; cAMP; EPAC; ERK1/2;