Biochimica et Biophysica Acta 1843 (2014) 398–435 Contents lists available at ScienceDirect Biochimica et Biophysica Acta journal homepage: www.elsevier.com/locate/bbamcr Review The many faces of calmodulin in cell proliferation, programmed cell death, autophagy, and cancer Martin W. Berchtold a,⁎, Antonio Villalobo b,⁎⁎ a Department of Biology, University of Copenhagen, Copenhagen Biocenter 4-2-09 Ole Maaløes Vej 5, DK-2200 Copenhagen N, Denmark b Instituto de Investigaciones Biomédicas, Department of Cancer Biology, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, c/Arturo Duperier 4, E-28029 Madrid, Spain article info abstract Article history: Calmodulin (CaM) is a ubiquitous Ca2+ receptor protein mediating a large number of signaling processes in all Received 14 September 2013 eukaryotic cells. CaM plays a central role in regulating a myriad of cellular functions via interaction with multiple Received in revised form 24 October 2013 target proteins. This review focuses on the action of CaM and CaM-dependent signaling systems in the control of Accepted 26 October 2013 vertebrate cell proliferation, programmed cell death and autophagy. The significance of CaM and interconnected Available online 2 November 2013 CaM-regulated systems for the physiology of cancer cells including tumor stem cells, and processes required for tumor progression such as growth, tumor-associated angiogenesis and metastasis are highlighted. Furthermore, Keywords: Calmodulin the potential targeting of CaM-dependent signaling processes for therapeutic use is discussed. Ca2+ binding protein © 2013 Elsevier B.V. All rights reserved. Cell proliferation Cancer biology Apoptosis Autophagy 1. Introduction the importance of this EF-hand type Ca2+-binding protein in cell phys- iology. Since its discovery as a cyclic nucleotide phosphodiesterase Calmodulin (CaM) is considered the major regulator of Ca2+- (PDE) activator in the early '70 of the past century [1,2],avastamount dependent signaling in all eukaryotic cells. Its ubiquitous distribution, of data has been accumulated on its important function in the control its high conservation during evolution, its broad spectrum of functions of numerous physiological processes such as e.g. cell motility, cytoskel- and the fact that it is essential in all so far tested organisms underlay eton architecture and function, cell proliferation, apoptosis, autophagy, metabolic homeostasis, phospholipid turnover, protein folding, phos- Abbreviations: B859-35, (−) enantiomer of dihydropyrine 3-methyl-5-3-(4,4-diphenyl-1- phorylation/dephosphorylation of proteins, ions transport, osmotic piperidinyl)-propyl-1,4-dihydro-2,6-dimethyl-4-(3-nitrophenyl)-piridine-3,5-dicarboxylate- control, reproductive processes, muscle contraction and gene expres- hydrochloride (niguldipine); CAPP1-CaM, norchlorpromazine-CaM adduct; DMBA, 7,12- sion among others. Much has been learned during the recent years on dimethylbenz[a]anthracene; EBB, O-(4-ethoxyl-butyl)-berbamine; FL-CaM, fluorescein-CaM fl the structure and functionality of CaM and its target proteins, the dy- adduct; FPCE, uphenazine-N-2-chloroethane; HBC, (4-[3,5-bis-[2-(4-hydroxy-3-methoxy- 2+ 2+ phenyl)-ethyl]-4,5-dihydro-pyrazol-1-yl]-benzoic acid; HBCP, (4-[3,5-bis-[2-(4-hydroxy- namics of Ca binding and the different Ca -dependent as well as 2+ 3-methoxy-phenyl)-vinyl]-4,5-dihydro-pyrazol-1-yl]-phenyl)-(4-methyl-piperazin-1-yl)- Ca -independent mechanisms of interaction with its multiple targets methanone; J-8, N-8-aminooctyl-5-iodo-naphthalenesulfonamide; KAR-2, 3′-(β-chloroethyl)- (reviewed in Refs. [3–8]). One reason for the unprecedented diversity ′ ′ ′ 2 ,4 -dioxo-3,5 -spiro-oxazolidino-4-deacetoxy-vinblastine; KN-62, 1-[N,O-bis(5- of CaM functions is connected with its ability to interact with several isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine; KN-93, N-[2-[N-(4- hundred different target proteins and to modulate their activity in chlorocinnamyl)-N-methylaminomethyl]phenyl]-N-(2-hydroxyethyl)-4-methoxybenzene- sulfonamide; RITC-CaM, rhodamine isothiocyanate-CaM adduct; TA-CaM, 2-chloro-(ε- many different ways. In this review we will describe accumulated infor- amino-Lys75)-[6-(4-(N,N′-diethylaminophenyl)-1,3,5-triazin-4-yl]-CaM adduct; TFP, trifluo- mation and recent advances on the action of CaM and different CaM- perazine; TPA, 12-O-tetradecanoyl-phorbol-13-acetate; W-5, N-(6-aminohexyl)-1- dependent systems in the control of cell proliferation, programmed naphthalenesulfonamide; W-7, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide; cell death and autophagy, with a particular focus on the implication of W-12, N-(4-aminobutyl)-2-naphthalenesulfonamide; W-13, N-(4-aminobutyl)-5-chloro-2- naphthalenesulfonamide these mechanisms for the physiology of tumor cells. 2+ ⁎ Correspondence to: M.W. Berchtold, Department of Biology, University of The transient increase in the concentration of free Ca in the cyto- Copenhagen, Copenhagen Biocenter 4-2-09 Ole Maaløes Vej 5, DK-2200 Copenhagen N, sol and its spread to the nucleus upon cell activation by a broad range of Denmark. Fax: +45 33935 220. stimuli including mitogenic factors and other agonists has been recog- ⁎⁎ Correspondence to: A. Villalobo, Instituto de Investigaciones Biomédicas, CSIC–UAM, nized as the principal event responsible for the initiation of many signal c/Arturo Duperier 4, E-28029 Madrid, Spain. Fax: +34 91 585 4401. E-mail addresses: [email protected] (M.W. Berchtold), transduction processes. The transport systems that control the concen- [email protected] (A. Villalobo). tration of Ca2+ in the nucleus are of upmost importance for cell 0167-4889/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.bbamcr.2013.10.021 M.W. Berchtold, A. Villalobo / Biochimica et Biophysica Acta 1843 (2014) 398–435 399 2+ proliferation, as Ca at this location not only binds to nuclear CaM but calcium-mobilizing messengers, e.g. IP3, across gap-junction channels, also controls the activity of many transcription factors (reviewed in Refs. each formed by six connexin molecules. Some connexins are CaM- [9,10]). The study of the complex network of Ca2+-binding proteins and binding proteins, e.g. connexin-32 and connexin-43 [58,59]. Alterna- their interactions, denoted as 'calciomics, try to understand the multiple tively, the release of ATP to the extracellular fluid across connexin hemi- effects of Ca2+ leading to changes in cell physiology (reviewed in Ref. channels and maxi-anion channels or by vesicular release may 2+ 2+ [11]). Besides CaM other Ca -binding proteins including e.g. calpains stimulate P2 receptors in neighboring cells where additional Ca sig- (reviewed in Ref. [12]), ALG-2 (apoptosis-linked gene 2 product) nals are elicited (reviewed in Ref. [60]). Moreover, the open/closed (reviewed in Ref. [13]), sorcin (reviewed in Ref. [14]), S-100 proteins states of connexin hemichannels appear to be controlled by both the ex- (reviewed in Ref. [15]), oncomodulin [16,17] and annexins (reviewed tracellular and intracellular concentrations of Ca2+ [61],andCa2+ can in Ref. [18]) also play important roles in signal transduction leading to itself permeate the hemichannels as demonstrated for example with the control of cell proliferation as well as cell death. However, this reconstituted connexin 26 in liposomes [62]. topic lies outside the scope of this review. The activation of Ca2+- The increase in intracellular Ca2+ triggers the formation of the Ca2+/ binding proteins occurs by Ca2+ entry into the cytosol through the CaM complex, as monitored in intact cells [63,64],followedbymodula- temporospatial opening of Ca2+ channels initiated by a variety of signal- tion of the functionality of the target proteins (Fig. 2). Saucermann and ing molecules. They may be derived from the metabolism of phospho- Bers [65] have proposed that there are two distinct pools of CaM in the lipids, such as inositol-1,4,5-trisphosphate (IP3)resultingfromthe cell: one, named ‘dedicated’, formed by CaM molecules already tethered hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase to their targets (e.g. Ca2+ channels) ready to activate them upon Ca2+ Cγ (PLCγ) (reviewed in Refs. [19–24]), and leukotriene C4 formed availability; and the other minor pool, ‘named promiscuous’ formed from arachidonic acid released by the action of phospholipase A2 by free CaM molecules in the cytosol, which may interact and regulate 2+ (PLA2) on phospholipids (reviewed in Refs. [25–27]). Additional mes- CaM-binding protein upon Ca binding (Fig. 2). Frequently, the inter- sengers implicated in Ca2+ mobilization are the nucleotides cyclic action of Ca2+/CaM with their targets implicate wrapping the CaM- ADP-ribose (cADPR) and nicotinic acid adenine dinucleotide phosphate binding region of the target by the CaM molecule collapsed by bending (NAADP) derived from NAD+ and NADP+, respectively. They activate around its central linker α-helix and bringing the N- and C-globular re- ryanodine receptors located at the endo(sarco)plasmic reticulum and gions closer to each other, as for example with CaM-dependent kinase two-pore channels located at endo-lysosomes, respectively (reviewed (CaMK)-I [66] (Fig. 3), CaMK-II [67] or calcineurin (CaN) also denoted in Refs. [28–31]). Another important Ca2+ mobilizing agent is ADP- protein phosphatase 2B (PP2B) [68]. CaMK-II, when activated by Ca2+ ribose (ADPR), synthesized by the poly(ADP-ribose) polymerase 1/2- and CaM converts