Abstract
Zyxin, a focal adhesion molecule, contains LIM domains and shuttles between the cytoplasm and the nucleus. Nuclear zyxin promotes cardiomyocyte survival, which is mediated by nuclear-activated Akt. However, the molecular mechanism of how zyxin antagonizes apoptosis remains elusive. Here, we report that zyxin binds to acinus-S, a nuclear speckle protein inducing apoptotic chromatin condensation after cleavage by caspases, and prevents its apoptotic action, which is regulated by Akt. Akt binds and phosphorylates zyxin on serine 142, leading to its association with acinus. Interestingly, 14-3-3γ, but not ζ isoform selectively, triggers zyxin nuclear translocation, which is Akt phosphorylation dependent. Zyxin is also a substrate of caspases, but Akt phosphorylation is unable to prevent its apoptotic cleavage. Expression of zyxin S142D, a phosphorylation mimetic mutant, diminishes acinus proteolytic cleavage and chromatin condensation; by contrast, wild-type zyxin or unphosphorylated S142A mutant fails. Thus, Akt regulates zyxin/acinus complex formation in the nucleus, contributing to suppression of apoptosis.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- aa:
-
amino acids
- ANP:
-
atrial natriuretic peptide
- ASAP:
-
apoptosis- and splicing-associated protein
- CA:
-
constitutively active
- CTD:
-
C-terminal domain
- EGF:
-
epidermal growth factor
- EJC:
-
exon junction complex
- KD:
-
kinase-dead
- MEF:
-
mouse embryonic fibroblast
- NES:
-
nuclear export signal
- NGF:
-
nerve growth factor
- NTD:
-
N-terminal domain
- PRD:
-
proline-rich domain
- STS:
-
staurosporine
References
Ahern-Djamali SM, Comer AR, Bachmann C, Kastenmeier AS, Reddy SK, Beckerle MC et al. Mutations in Drosophila enabled and rescue by human vasodilator-stimulated phosphoprotein (VASP) indicate important functional roles for Ena/VASP homology domain 1 (EVH1) and EVH2 domains. Mol Biol Cell 1998; 9: 2157–2171.
Crawford AW, Michelsen JW, Beckerle MC . An interaction between zyxin and alpha-actinin. J Cell Biol 1992; 116: 1381–1393.
Drees BE, Andrews KM, Beckerle MC . Molecular dissection of zyxin function reveals its involvement in cell motility. J Cell Biol 1999; 147: 1549–1560.
Gertler FB, Niebuhr K, Reinhard M, Wehland J, Soriano P . Mena, a relative of VASP and Drosophila Enabled, is implicated in the control of microfilament dynamics. Cell 1996; 87: 227–239.
Niebuhr K, Ebel F, Frank R, Reinhard M, Domann E, Carl UD et al. A novel proline-rich motif present in ActA of Listeria monocytogenes and cytoskeletal proteins is the ligand for the EVH1 domain, a protein module present in the Ena/VASP family. EMBO J 1997; 16: 5433–5444.
Reinhard M, Zumbrunn J, Jaquemar D, Kuhn M, Walter U, Trueb B . An alpha-actinin binding site of zyxin is essential for subcellular zyxin localization and alpha-actinin recruitment. J Biol Chem 1999; 274: 13410–13418.
Hobert O, Schilling JW, Beckerle MC, Ullrich A, Jallal B . SH3 domain-dependent interaction of the proto-oncogene product Vav with the focal contact protein zyxin. Oncogene 1996; 12: 1577–1581.
Gill GN . The enigma of LIM domains. Structure 1995; 3: 1285–1289.
Sanchez-Garcia I, Rabbitts TH . The LIM domain: a new structural motif found in zinc-finger-like proteins. Trends Genet 1994; 10: 315–320.
Nix DA, Beckerle MC . Nuclear–cytoplasmic shuttling of the focal contact protein, zyxin: a potential mechanism for communication between sites of cell adhesion and the nucleus. J Cell Biol 1997; 138: 1139–1147.
Missero C, Pirro MT, Simeone S, Pischetola M, Di Lauro R . The DNA glycosylase T:G mismatch-specific thymine DNA glycosylase represses thyroid transcription factor-1-activated transcription. J Biol Chem 2001; 276: 33569–33575.
Sahara S, Aoto M, Eguchi Y, Imamoto N, Yoneda Y, Tsujimoto Y . Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation. Nature 1999; 401: 168–173.
Rappsilber J, Ryder U, Lamond AI, Mann M . Large-scale proteomic analysis of the human spliceosome. Genome Res 2002; 12: 1231–1245.
Schwerk C, Prasad J, Degenhardt K, Erdjument-Bromage H, White E, Tempst P et al. ASAP, a novel protein complex involved in RNA processing and apoptosis. Mol Cell Biol 2003; 23: 2981–2990.
Joselin AP, Schulze-Osthoff K, Schwerk C . Loss of Acinus inhibits oligonucleosomal DNA fragmentation but not chromatin condensation during apoptosis. J Biol Chem 2006; 281: 12475–12484.
Tange TO, Shibuya T, Jurica MS, Moore MJ . Biochemical analysis of the EJC reveals two new factors and a stable tetrameric protein core. RNA 2005; 11: 1869–1883.
Hu Y, Yao J, Liu Z, Liu X, Fu H, Ye K . Akt phosphorylates acinus and inhibits its proteolytic cleavage, preventing chromatin condensation. EMBO J 2005; 24: 3543–3554.
Kato T, Muraski J, Chen Y, Tsujita Y, Wall J, Glembotski CC et al. Atrial natriuretic peptide promotes cardiomyocyte survival by cGMP-dependent nuclear accumulation of zyxin and Akt. J Clin Invest 2005; 115: 2716–2730.
Bavelloni A, Faenza I, Cioffi G, Piazzi M, Parisi D, Matic I et al. Proteomic-based analysis of nuclear signaling: PLCβ1 affects the expression of the splicing factor SRp20 in Friend erythroleukemia cells. Proteomics 2006; 6: 5725–5734.
Borgatti P, Martelli AM, Tabellini G, Bellacosa A, Capitani S, Neri LM . Threonine 308 phosphorylated form of Akt translocates to the nucleus of PC12 cells under nerve growth factor stimulation and associates with the nuclear matrix protein nucleolin. J Cell Physiol 2003; 196: 79–88.
Xuan Nguyen TL, Choi JW, Lee SB, Ye K, Woo SD, Lee KH et al. Akt phosphorylation is essential for nuclear translocation and retention in NGF-stimulated PC12 cells. Biochem Biophys Res Commun 2006; 349: 789–798.
Fu H, Subramanian RR, Masters SC . 14-3-3 proteins: structure, function, and regulation. Annu Rev Pharmacol Toxicol 2000; 40: 617–647.
Brunet A, Kanai F, Stehn J, Xu J, Sarbassova D, Frangioni JV et al. 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. J Cell Biol 2002; 156: 817–828.
Liu X, Kim CN, Yang J, Jemmerson R, Wang X . Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell 1996; 86: 147–157.
Nix DA, Fradelizi J, Bockholt S, Menichi B, Louvard D, Friederich E et al. Targeting of zyxin to sites of actin membrane interaction and to the nucleus. J Biol Chem 2001; 276: 34759–34767.
Faul C, Huttelmaier S, Oh J, Hachet V, Singer RH, Mundel P . Promotion of importin alpha-mediated nuclear import by the phosphorylation-dependent binding of cargo protein to 14-3-3. J Cell Biol 2005; 169: 415–424.
Ahn JY, Rong R, Kroll TG, Van Meir EG, Snyder SH, Ye K . PIKE (phosphatidylinositol 3-kinase enhancer)-A GTPase stimulates Akt activity and mediates cellular invasion. J Biol Chem 2004; 279: 16441–16451.
Ye K, Compton DA, Lai MM, Walensky LD, Snyder SH . Protein 4.1N binding to nuclear mitotic apparatus protein in PC12 cells mediates the antiproliferative actions of nerve growth factor. J Neurosci 1999; 19: 10747–10756.
Ye K, Ke Y, Keshava N, Shanks J, Kapp JA, Tekmal RR, Petros J, Joshi HC . Opium alkaloid noscapine is an antitumor agent that arrests metaphase and induces apoptosis in dividing cells. Proc Natl Acad Sci USA 1998; 95: 1601–1606.
Acknowledgements
This work was supported by grants from National Institute of Health (RO1, NS045627, CA117872) to K Ye. We are thankful to Dr Mark Sussman for NLS-Akt and GFP-NLS-zyxin adenovirus and Dr Jürgen Wehland for the GFP-zyxin construct.
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by P Mehlen
Rights and permissions
About this article
Cite this article
Chan, CB., Liu, X., Tang, X. et al. Akt phosphorylation of zyxin mediates its interaction with acinus-S and prevents acinus-triggered chromatin condensation. Cell Death Differ 14, 1688–1699 (2007). https://doi.org/10.1038/sj.cdd.4402179
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4402179
Keywords
This article is cited by
-
Cell density-dependent proteolysis by HtrA1 induces translocation of zyxin to the nucleus and increased cell survival
Cell Death & Disease (2020)
-
Zyxin is a novel interacting partner for SIRT1
BMC Cell Biology (2009)
-
NGF inhibits human leukemia proliferation by downregulating cyclin A1 expression through promoting acinus/CtBP2 association
Oncogene (2009)
