Molecular mechanisms underlying nucleocytoplasmic shuttling of actinin-4

In addition to its well-known role as a crosslinker of actin filaments at focal-adhesion sites, actinin-4 is known to be localized to the nucleus. In this study, we reveal the molecular mechanism underlying nuclear localization of actinin-4 and its novel interactions with transcriptional regulators. We found that actinin-4 is imported into the nucleus through the nuclear pore complex in an importin-independent manner and is exported by the chromosome region maintenance-1 (CRM1)-dependent pathway. Nuclear actinin-4 levels were significantly increased in the late G2 phase of the cell cycle and were decreased in the G1 phase, suggesting that active release from the actin cytoskeleton was responsible for increased nuclear actinin-4 in late G2. Nuclear actinin-4 was found to interact with the INO80 chromatin-remodeling complex. It also directs the expression of a subset of cell-cycle-related genes and interacts with the upstream-binding factor (UBF)-dependent rRNA transcriptional machinery in the M phase. These findings provide molecular mechanisms for both nucleocytoplasmic shuttling of proteins that do not contain a nuclear-localization signal and cell-cycle-dependent gene regulation that reflects morphological changes in the cytoskeleton.

[1]  H. Sweeney,et al.  Sarcomeric-alpha-actinin defective in vinculin-binding causes Z-line expansion and nemaline-like body formation in cultured chick myotubes. , 2009, Experimental cell research.

[2]  K. Takeyasu,et al.  Proteomic and targeted analytical identification of BXDC1 and EBNA1BP2 as dynamic scaffold proteins in the nucleolus , 2009, Genes to cells : devoted to molecular & cellular mechanisms.

[3]  T. Uchida,et al.  The human actin-related protein hArp5: nucleo-cytoplasmic shuttling and involvement in DNA repair. , 2009, Experimental cell research.

[4]  S. Otsuka,et al.  Individual binding pockets of importin-β for FG-nucleoporins have different binding properties and different sensitivities to RanGTP , 2008, Proceedings of the National Academy of Sciences.

[5]  K. Takeyasu,et al.  Nuclear matrix contains novel WD‐repeat and disordered‐region‐rich proteins , 2008, FEBS letters.

[6]  S. Lim,et al.  FAK nuclear export signal sequences , 2008, FEBS letters.

[7]  K. Djinović-Carugo,et al.  α-Actinin structure and regulation , 2008, Cellular and Molecular Life Sciences.

[8]  H. Szerlong,et al.  The HSA domain binds nuclear actin-related proteins to regulate chromatin-remodeling ATPases , 2008, Nature Structural &Molecular Biology.

[9]  S. Gasser,et al.  Ino80 Chromatin Remodeling Complex Promotes Recovery of Stalled Replication Forks , 2008, Current Biology.

[10]  K. Magnusson,et al.  RelA/NF-kappaB transcription factor associates with alpha-actinin-4. , 2008, Experimental cell research.

[11]  M. Washburn,et al.  YY1 functions with INO80 to activate transcription , 2007, Nature Structural &Molecular Biology.

[12]  M. Rexach,et al.  Natively Unfolded Nucleoporins Gate Protein Diffusion across the Nuclear Pore Complex , 2007, Cell.

[13]  Xiaofan Li,et al.  α-Actinin 4 Potentiates Myocyte Enhancer Factor-2 Transcription Activity by Antagonizing Histone Deacetylase 7* , 2006, Journal of Biological Chemistry.

[14]  Daniel Nietlispach,et al.  The Vinculin Binding Sites of Talin and α-Actinin Are Sufficient to Activate Vinculin* , 2006, Journal of Biological Chemistry.

[15]  Michael K. Coleman,et al.  A Mammalian Chromatin Remodeling Complex with Similarities to the Yeast INO80 Complex* , 2005, Journal of Biological Chemistry.

[16]  Clemens Vonrhein,et al.  Structural Dynamics of α-Actinin-Vinculin Interactions , 2005, Molecular and Cellular Biology.

[17]  K. Fukui,et al.  Proteome Analysis of Human Metaphase Chromosomes* , 2005, Journal of Biological Chemistry.

[18]  K. G. Young,et al.  Spectrin repeat proteins in the nucleus , 2005, BioEssays : news and reviews in molecular, cellular and developmental biology.

[19]  Barbara Hohn,et al.  Recruitment of the INO80 Complex by H2A Phosphorylation Links ATP-Dependent Chromatin Remodeling with DNA Double-Strand Break Repair , 2004, Cell.

[20]  Si Wu,et al.  A role for beta-actin in RNA polymerase III transcription. , 2004, Genes & development.

[21]  H. Zentgraf,et al.  Nuclear actin and myosin I are required for RNA polymerase I transcription , 2004, Nature Cell Biology.

[22]  T. Hope,et al.  Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II , 2004, Nature Cell Biology.

[23]  O. Carpén,et al.  Alpha-actinin revisited: a fresh look at an old player. , 2004, Cell motility and the cytoskeleton.

[24]  L. Backman,et al.  Molecular Evolution and Structure of α-Actinin , 2004 .

[25]  K. G. Young,et al.  Bpag1 localization to actin filaments and to the nucleus is regulated by its N-terminus , 2003, Journal of Cell Science.

[26]  S. Yoshimura,et al.  On-substrate lysis treatment combined with scanning probe microscopy revealed chromosome structures in eukaryotes and prokaryotes. , 2003, Journal of electron microscopy.

[27]  Carl Wu,et al.  Involvement of actin-related proteins in ATP-dependent chromatin remodeling. , 2003, Molecular cell.

[28]  I. Zachary,et al.  The focal adhesion kinase amino-terminal domain localises to nuclei and intercellular junctions in HEK 293 and MDCK cells independently of tyrosine 397 and the carboxy-terminal domain. , 2002, Biochemical and biophysical research communications.

[29]  Denis Hochstrasser,et al.  Functional proteomic analysis of human nucleolus. , 2002, Molecular biology of the cell.

[30]  M. Mann,et al.  Directed Proteomic Analysis of the Human Nucleolus , 2002, Current Biology.

[31]  M. Saraste,et al.  Crystal Structure of the α-Actinin Rod Reveals an Extensive Torsional Twist , 2001 .

[32]  I. Zachary,et al.  Nuclear localization and apoptotic regulation of an amino-terminal domain focal adhesion kinase fragment in endothelial cells. , 2000, Biochemical and biophysical research communications.

[33]  D. Stillman,et al.  Multiple actin-related proteins of Saccharomyces cerevisiae are present in the nucleus. , 2000, Journal of biochemistry.

[34]  S. Hirohashi,et al.  Actinin-4 is preferentially involved in circular ruffling and macropinocytosis in mouse macrophages: analysis by fluorescence ratio imaging. , 2000, Journal of cell science.

[35]  A. Evans,et al.  The human non-muscle α-actinin protein encoded by the ACTN4 gene suppresses tumorigenicity of human neuroblastoma cells , 2000, Oncogene.

[36]  Hitoshi Tsuda,et al.  Actinin-4, a Novel Actin-bundling Protein Associated with Cell Motility and Cancer Invasion , 1998, The Journal of cell biology.

[37]  M. Yanagida,et al.  Molecular Cloning and Cell Cycle-dependent Expression of Mammalian CRM1, a Protein Involved in Nuclear Export of Proteins* , 1997, The Journal of Biological Chemistry.

[38]  K. Hirota,et al.  Mitosis specific serine phosphorylation and downregulation of one of the focal adhesion protein, paxillin , 1997, Oncogene.

[39]  F. Bischoff,et al.  Dominant‐negative mutants of importin‐β block multiple pathways of import and export through the nuclear pore complex , 1997, The EMBO journal.

[40]  F. Bischoff,et al.  Identification of different roles for RanGDP and RanGTP in nuclear protein import. , 1996, The EMBO journal.

[41]  B. Cullen,et al.  Protein sequence requirements for function of the human T-cell leukemia virus type 1 Rex nuclear export signal delineated by a novel in vivo randomization-selection assay , 1996, Molecular and cellular biology.

[42]  D. Hernandez-Verdun,et al.  The rDNA transcription machinery is assembled during mitosis in active NORs and absent in inactive NORs , 1996, The Journal of cell biology.

[43]  E. Hartmann,et al.  Distinct functions for the two importin subunits in nuclear protein import , 1995, Nature.

[44]  A. Blanchard,et al.  Identification of the vinculin-binding site in the cytoskeletal protein α-actinin , 1994 .

[45]  T. Masaki,et al.  Molecular cloning of low-Ca2+-sensitive-type non-muscle α-actinin , 1994 .

[46]  T. Byers,et al.  Cloning and characterization of two human skeletal muscle alpha-actinin genes located on chromosomes 1 and 11. , 1992, The Journal of biological chemistry.

[47]  D. Kwiatkowski,et al.  Cloning and chromosomal localization of the human cytoskeletal alpha-actinin gene reveals linkage to the beta-spectrin gene. , 1990, American journal of human genetics.

[48]  A. Blanchard,et al.  The cDNA sequence of a human placental α-actinin , 1989 .

[49]  M. Davison,et al.  α-actinins and the DMD protein contain spectrin-like repeats , 1988, Cell.

[50]  A. Asano,et al.  Further characterization of a conserved actin-binding 27-kDa fragment of actinogelin and alpha-actinins and mapping of their binding sites on the actin molecule by chemical cross-linking. , 1987, The Journal of biological chemistry.

[51]  S. Penman,et al.  The nonchromatin substructures of the nucleus: the ribonucleoprotein (RNP)-containing and RNP-depleted matrices analyzed by sequential fractionation and resinless section electron microscopy , 1986, The Journal of cell biology.

[52]  D. Nietlispach,et al.  The vinculin binding sites of talin and alpha-actinin are sufficient to activate vinculin. , 2006, The Journal of biological chemistry.

[53]  Clemens Vonrhein,et al.  Structural dynamics of alpha-actinin-vinculin interactions. , 2005, Molecular and cellular biology.

[54]  L. Backman,et al.  Molecular evolution and structure of alpha-actinin. , 2004, Molecular biology and evolution.

[55]  Søren Brunak,et al.  NESbase version 1.0: a database of nuclear export signals , 2003, Nucleic Acids Res..

[56]  M. Saraste,et al.  Crystal structure of the alpha-actinin rod reveals an extensive torsional twist. , 2001, Structure.

[57]  A. Blanchard,et al.  Identification of the vinculin-binding site in the cytoskeletal protein alpha-actinin. , 1994, The Biochemical journal.

[58]  T. Masaki,et al.  Molecular cloning of low-Ca(2+)-sensitive-type non-muscle alpha-actinin. , 1994, European journal of biochemistry.

[59]  A. Blanchard,et al.  The cDNA sequence of a human placental alpha-actinin. , 1989, Nucleic acids research.