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Ultrastructural localization of actin and actin-binding proteins in the nucleus

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Abstract

Nuclear actin plays an important role in such processes as chromatin remodeling, transcriptional regulation, RNA processing, and nuclear export. Recent research has demonstrated that actin in the nucleus probably exists in dynamic equilibrium between monomeric and polymeric forms, and some of the actin-binding proteins, known to regulate actin dynamics in cytoplasm, have been also shown to be present in the nucleus. In this paper, we present ultrastructural data on distribution of actin and various actin-binding proteins (α-actinin, filamin, p190RhoGAP, paxillin, spectrin, and tropomyosin) in nuclei of HeLa cells and resting human lymphocytes. Probing extracts of HeLa cells for the presence of actin-binding proteins also confirmed their presence in nuclei. We report for the first time the presence of tropomyosin and p190RhoGAP in the cell nucleus, and the spatial colocalization of actin with spectrin, paxillin, and α-actinin in the nucleolus.

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References

  • Babakov VN, Bobkov DE, Petukhova OA, Turoverova LV, Kropacheva IV, Podol’skaia EP, Pinaev GP (2004) alpha-Actinin-4 and p65/RelA subunit of NF-kappaB transcription factor are co-localized and migrate together into the nucleus in EGF-stimulated A431 cell. Tsitologiia 46:1064–1072

    PubMed  CAS  Google Scholar 

  • Bachs O, Lanini L, Serratosa J, Coll MJ, Bastos R, Aligue R, Rius E, Carafoli E (1990) Calmodulin-binding proteins in the nuclei of quiescent and proliferatively activated rat liver cells. J Biol Chem 265:18595–18600

    PubMed  CAS  Google Scholar 

  • Bennett V (1990) Spectrin-based membrane skeleton: a multipotential adaptor between plasma membrane and cytoplasm. Physiol Rev 70:1029–1065

    PubMed  CAS  Google Scholar 

  • Boyum A (1968) A one-stage procedure for isolation of granulocytes and lymphocytes from human blood. General sedimentation properties of white blood cells in a 1 g gravity field. Scand J Clin Lab Invest Suppl 97:51–76

    PubMed  CAS  Google Scholar 

  • Burridge K, Kelly T, Mangeat P (1982) Nonerythrocyte spectrins: actin-membrane attachment proteins occurring in many cell types. J Cell Biol 95:478–486

    Article  PubMed  CAS  Google Scholar 

  • Chakraborty S, Reineke EL, Lam M, Li X, Liu Y, Gao C, Khurana S, Kao HY (2006) Alpha-actinin 4 potentiates myocyte enhancer factor-2 transcription activity by antagonizing histone deacetylase 7. J Biol Chem 281:35070–35080

    Article  PubMed  CAS  Google Scholar 

  • Chang JH, Gill S, Settleman J, Parsons SJ (1995) c-Src regulates the simultaneous rearrangement of actin cytoskeleton, p190RhoGAP, and p120RasGAP following epidermal growth factor stimulation. J Cell Biol 130:355–368

    Article  PubMed  CAS  Google Scholar 

  • Chuang CH, Carpenter AE, Fuchsova B, Johnson T, de Lanerolle P, Belmont AS (2006) Long-range directional movement of an interphase chromosome site. Curr Biol 16:825–831

    Article  PubMed  CAS  Google Scholar 

  • de Lanerolle P, Johnson T, Hofmann WA (2005) Actin and myosin I in the nucleus: what next? Nat Struct Mol Biol 12:742–746

    Article  PubMed  Google Scholar 

  • Djinovic-Carugo K, Gautel M, Ylanne J, Young P (2002) The spectrin repeat: a structural platform for cytoskeletal protein assemblies. FEBS Lett 513:119–123

    Article  PubMed  CAS  Google Scholar 

  • Egly JM, Miyamoto NG, Moncollin V, Chambon P (1984) Is actin a transcription initiation factor for RNA polymerase B? EMBO J 3:2363–2371

    PubMed  CAS  Google Scholar 

  • Fomproix N, Percipalle P (2004) An actin-myosin complex on actively transcribing genes. Exp Cell Res 294:140–148

    Article  PubMed  CAS  Google Scholar 

  • Forest T, Barnard S, Baines JD (2005) Active intranuclear movement of herpesvirus capsids. Nat Cell Biol 7:429–431

    Article  PubMed  CAS  Google Scholar 

  • Gettemans J, Van Impe K, Delanote V, Hubert T, Vandekerckhove J, De Corte V (2005) Nuclear actin-binding proteins as modulators of gene transcription. Traffic 6:847–857

    Article  PubMed  CAS  Google Scholar 

  • Glenney JR Jr, Glenney P (1983) Fodrin is the general spectrin-like protein found in most cells whereas spectrin and the TW protein have a restricted distribution. Cell 34:503–512

    Article  PubMed  CAS  Google Scholar 

  • Gonsior SM, Platz S, Buchmeier S, Scheer U, Jockusch BM, Hinssen H (1999) Conformational difference between nuclear and cytoplasmic actin as detected by a monoclonal antibody. J Cell Sci 112(Pt 6):797–809

    PubMed  CAS  Google Scholar 

  • Grummt I (2006) Actin and myosin as transcription factors. Curr Opin Genet Dev 16:191–196

    Article  PubMed  CAS  Google Scholar 

  • Higaki T, Sano T, Hasezawa S (2007) Actin microfilament dynamics and actin side-binding proteins in plants. Curr Opin Plant Biol 10:549–556

    Article  PubMed  CAS  Google Scholar 

  • Hofmann W, Reichart B, Ewald A, Muller E, Schmitt I, Stauber RH, Lottspeich F, Jockusch BM, Scheer U, Hauber J, Dabauvalle MC (2001) Cofactor requirements for nuclear export of Rev response element (RRE)- and constitutive transport element (CTE)-containing retroviral RNAs. An unexpected role for actin. J Cell Biol 152:895–910

    Article  PubMed  CAS  Google Scholar 

  • Holaska JM, Kowalski AK, Wilson KL (2004) Emerin caps the pointed end of actin filaments: evidence for an actin cortical network at the nuclear inner membrane. PLoS Biol 2:E231

    Article  PubMed  Google Scholar 

  • Honda K, Yamada T, Endo R, Ino Y, Gotoh M, Tsuda H, Yamada Y, Chiba H, Hirohashi S (1998) Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion. J Cell Biol 140:1383–1393

    Article  PubMed  CAS  Google Scholar 

  • Hozák P, Cook PR, Schöfer C, Mosgoller W, Wachtler F (1994) Site of transcription of ribosomal RNA and intranucleolar structure in HeLa cells. J Cell Sci 107:639–648

    PubMed  Google Scholar 

  • Hu P, Wu S, Hernandez N (2004) A role for beta-actin in RNA polymerase III transcription. Genes Dev 18:3010–3015

    Article  PubMed  CAS  Google Scholar 

  • Jockusch BM, Schoenenberger CA, Stetefeld J, Aebi U (2006) Tracking down the different forms of nuclear actin. Trends Cell Biol 16:391–396

    Article  PubMed  CAS  Google Scholar 

  • Krauss SW, Chen C, Penman S, Heald R (2003) Nuclear actin and protein 4.1: essential interactions during nuclear assembly in vitro. Proc Natl Acad Sci U S A 100:10752–10757

    Article  PubMed  CAS  Google Scholar 

  • Kukalev A, Nord Y, Palmberg C, Bergman T, Percipalle P (2005) Actin and hnRNP U cooperate for productive transcription by RNA polymerase II. Nat Struct Mol Biol 12:238–244

    Article  PubMed  CAS  Google Scholar 

  • Lin JJ (1981) Monoclonal antibodies against myofibrillar components of rat skeletal muscle decorate the intermediate filaments of cultured cells. Proc Natl Acad Sci U S A 78:2335–2339

    Article  PubMed  CAS  Google Scholar 

  • Loy CJ, Sim KS, Yong EL (2003) Filamin-A fragment localizes to the nucleus to regulate androgen receptor and coactivator functions. Proc Natl Acad Sci U S A 100:4562–4567

    Article  PubMed  CAS  Google Scholar 

  • Lv L, Xu J, Zhao S, Chen C, Zhao X, Gu S, Ji C, Xie Y, Mao Y (2007) Sequence analysis of a human RhoGAP domain-containing gene and characterization of its expression in human multiple tissues. DNA Seq 18:184–189

    Article  PubMed  CAS  Google Scholar 

  • Machesky LM, Mullins RD, Higgs HN, Kaiser DA, Blanchoin L, May RC (1999) Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex. Proc Natl Acad Sci U S A 96(7):3739–3744

    Article  PubMed  CAS  Google Scholar 

  • Marchesi VT, Steers E Jr (1968) Selective solubilization of a protein component of the red cell membrane. Science 159:203–204

    Article  PubMed  CAS  Google Scholar 

  • McDonald D, Carrero G, Andrin C, de Vries G, Hendzel MJ (2006) Nucleoplasmic beta-actin exists in a dynamic equilibrium between low-mobility polymeric species and rapidly diffusing populations. J Cell Biol 172:541–552

    Article  PubMed  CAS  Google Scholar 

  • McMahon LW, Sangerman J, Goodman SR, Kumaresan K, Lambert MW (2001) Human alpha spectrin II and the FANCA, FANCC, and FANCG proteins bind to DNA containing psoralen interstrand cross-links. Biochemistry 40:7025–7034

    Article  PubMed  CAS  Google Scholar 

  • Meng X, Yuan Y, Maestas A, Shen Z (2004) Recovery from DNA damage-induced G2 arrest requires actin-binding protein filamin-A/actin-binding protein 280. J Biol Chem 279:6098–6105

    Article  PubMed  CAS  Google Scholar 

  • Meyer RK, Aebi U (1990) Bundling of actin filaments by alpha-actinin depends on its molecular length. J Cell Biol 110:2013–2024

    Article  PubMed  CAS  Google Scholar 

  • Ogawa M, Hiraoka Y, Taniguchi K, Sakai Y, Aiso S (2001) mRNA sequence of the Xenopus laevis paxillin gene and its expression. Biochim Biophys Acta 1519:235–240

    PubMed  CAS  Google Scholar 

  • Ogawa M, Hiraoka Y, Aiso S (2003) Nuclear translocation of Xenopus laevis paxillin. Biochem Biophys Res Commun 304:676–683

    Article  PubMed  CAS  Google Scholar 

  • Olave IA, Reck-Peterson SL, Crabtree GR (2002) Nuclear actin and actin-related proteins in chromatin remodeling. Annu Rev Biochem 71:755–781

    Article  PubMed  CAS  Google Scholar 

  • Otey CA, Carpen O (2004) Alpha-actinin revisited: a fresh look at an old player. Cell Motil Cytoskeleton 58:104–111

    Article  PubMed  CAS  Google Scholar 

  • Ozanne DM, Brady ME, Cook S, Gaughan L, Neal DE, Robson CN (2000) Androgen receptor nuclear translocation is facilitated by the f-actin cross-linking protein filamin. Mol Endocrinol 14:1618–1626

    Article  PubMed  CAS  Google Scholar 

  • Pak CW, Flynn KC, Bamburg JR (2008) Actin-binding proteins take the reins in growth cones. Nat Rev Neurosci 9:136–147

    Article  PubMed  CAS  Google Scholar 

  • Pederson T (2008) As functional nuclear actin comes into view, is it globular, filamentous, or both? J Cell Biol 180(6):1061–1064

    Article  PubMed  CAS  Google Scholar 

  • Pederson T, Aebi U (2005) Nuclear actin extends, with no contraction in sight. Mol Biol Cell 16:5055–5060

    Article  PubMed  CAS  Google Scholar 

  • Pelletier O, Pokidysheva E, Hirst LS, Bouxsein N, Li Y, Safinya CR (2003) Structure of actin cross-linked with alpha-actinin: a network of bundles. Phys Rev Lett 91:148102

    Article  PubMed  CAS  Google Scholar 

  • Percipalle P, Visa N (2006) Molecular functions of nuclear actin in transcription. J Cell Biol 172:967–971

    Article  PubMed  CAS  Google Scholar 

  • Percipalle P, Fomproix N, Kylberg K, Miralles F, Bjorkroth B, Daneholt B, Visa N (2003) An actin-ribonucleoprotein interaction is involved in transcription by RNA polymerase II. Proc Natl Acad Sci U S A 100:6475–6480

    Article  PubMed  CAS  Google Scholar 

  • Percipalle P, Fomproix N, Cavellan E, Voit R, Reimer G, Kruger T, Thyberg J, Scheer U, Grummt I, Farrants AK (2006) The chromatin remodelling complex WSTF-SNF2 h interacts with nuclear myosin 1 and has a role in RNA polymerase I transcription. EMBO Rep 7:525–530

    PubMed  CAS  Google Scholar 

  • Philimonenko AA, Janáček J, Hozák P (2000) Statistical evaluation of colocalization patterns in immunogold labeling experiments. J Struct Biol 132:201–210

    Article  PubMed  CAS  Google Scholar 

  • Philimonenko VV, Zhao J, Iben S, Dingova H, Kysela K, Kahle M, Zentgraf H, Hofmann WA, de Lanerolle P, Hozák P, Grummt I (2004) Nuclear actin and myosin I are required for RNA polymerase I transcription. Nat Cell Biol 6:1165–1172

    Article  PubMed  CAS  Google Scholar 

  • Poch MT, Al-Kassim L, Smolinski SM, Hines RN (2004) Two distinct classes of CCAAT box elements that bind nuclear factor-Y/alpha-actinin-4: potential role in human CYP1A1 regulation. Toxicol Appl Pharmacol 199:239–250

    Article  PubMed  CAS  Google Scholar 

  • Pollard TD (2007) Regulation of actin filament assembly by Arp2/3 complex and formins. Annu Rev Biophys Biomol Struct 36:451–477

    Article  PubMed  CAS  Google Scholar 

  • Rando OJ, Zhao K, Crabtree GR (2000) Searching for a function for nuclear actin. Trends Cell Biol 10:92–97

    Article  PubMed  CAS  Google Scholar 

  • Repasky EA, Granger BL, Lazarides E (1982) Widespread occurrence of avian spectrin in nonerythroid cells. Cell 29:821–833

    Article  PubMed  CAS  Google Scholar 

  • Rohatgi R, Ma L, Miki H, Lopez M, Kirchhausen T, Takenawa T, Kirschner MW (1999) The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly. Cell 97:221–331

    Article  PubMed  CAS  Google Scholar 

  • Sagot I, Rodal AA, Moseley J, Goode BL, Pellman D (2002) An actin nucleation mechanism mediated by Bni1 and profilin. Nat Cell Biol 4(8):626–631

    PubMed  CAS  Google Scholar 

  • Schafer DA, Cooper JA (1995) Control of actin assembly at filament ends. Annu Rev Cell Dev Biol 11:497–518

    Article  PubMed  CAS  Google Scholar 

  • Schaller MD (2004) FAK and paxillin: regulators of N-cadherin adhesion and inhibitors of cell migration? J Cell Biol 166:157–159

    Article  PubMed  CAS  Google Scholar 

  • Scheer U, Hinssen H, Franke WW, Jockusch BM (1984) Microinjection of actin-binding proteins and actin antibodies demonstrates involvement of nuclear actin in transcription of lampbrush chromosomes. Cell 39:111–122

    Article  PubMed  CAS  Google Scholar 

  • Schleicher M, Jockusch BM (2008) Actin: its cumbersome pilgrimage through cellular compartments. Histochem Cell Biol 129:695–704

    Article  PubMed  CAS  Google Scholar 

  • Schoenenberger CA, Buchmeier S, Boerries M, Sutterlin R, Aebi U, Jockusch BM (2005) Conformation-specific antibodies reveal distinct actin structures in the nucleus and the cytoplasm. J Struct Biol 152:157–168

    Article  PubMed  CAS  Google Scholar 

  • Schöfer C, Janáček J, Weipoltshammer K, Pourani J, Hozák P (2004) Mapping of cellular compartments based on ultrastructural immunogold labeling. J Struct Biol 147:128–135

    Article  PubMed  Google Scholar 

  • Schwarzacher HG, Mikelsaar AV, Schnedl W (1978) The nature of the Ag-staining of nucleolus organizer regions. Electron- and light-microscopic studies on human cells in interphase, mitosis, and meiosis. Cytogenet Cell Genet 20:24–39

    Article  PubMed  CAS  Google Scholar 

  • Smith SS, Kelly KH, Jockusch BM (1979) Actin co-purifies with RNA polymerase II. Biochem Biophys Res Commun 86:161–166

    Article  PubMed  CAS  Google Scholar 

  • Sridharan D, Brown M, Lambert WC, McMahon LW, Lambert MW (2003) Nonerythroid alphaII spectrin is required for recruitment of FANCA and XPF to nuclear foci induced by DNA interstrand cross-links. J Cell Sci 116:823–835

    Article  PubMed  CAS  Google Scholar 

  • Sridharan DM, McMahon LW, Lambert MW (2006) alphaII-Spectrin interacts with five groups of functionally important proteins in the nucleus. Cell Biol Int 30:866–878

    Article  PubMed  CAS  Google Scholar 

  • Stuven T, Hartmann E, Gorlich D (2003) Exportin 6: a novel nuclear export receptor that is specific for profilin-actin complexes. EMBO J 22:5928–5940

    Article  PubMed  Google Scholar 

  • Tse WT, Tang J, Jin O, Korsgren C, John KM, Kung AL, Gwynn B, Peters LL, Lux SE (2001) A new spectrin, beta IV, has a major truncated isoform that associates with promyelocytic leukemia protein nuclear bodies and the nuclear matrix. J Biol Chem 276:23974–23985

    Article  PubMed  CAS  Google Scholar 

  • Uribe R, Jay D (2007) A review of actin binding proteins: new perspectives. Mol Biol Rep (Epub ahead of print). doi:10.1007/s11033-007-9159-2

  • van der Flier A, Sonnenberg A (2001) Structural and functional aspects of filamins. Biochim Biophys Acta 1538:99–117

    Article  PubMed  Google Scholar 

  • Vartiainen MK, Guettler S, Larijani B, Treisman R (2007) Nuclear actin regulates dynamic subcellular localization and activity of the SRF cofactor MAL. Science 316:1749–1752

    Article  PubMed  CAS  Google Scholar 

  • Vendrell M, Aligue R, Bachs O, Seratosa J (1991) Presence of calmodulin and calmodulin-binding proteins in the nuclei of brain cells. J Neurochem 57:622–628

    Article  PubMed  CAS  Google Scholar 

  • Vrhovski B, Schevzov G, Dingle S, Lessard JL, Gunning P, Weinberger RP (2003) Tropomyosin isoforms from the gamma gene differing at the C-terminus are spatially and developmentally regulated in the brain. J Neurosci Res 72:373–383

    Article  PubMed  CAS  Google Scholar 

  • Woods AJ, Roberts MS, Choudhary J, Barry ST, Mazaki Y, Sabe H, Morley SJ, Critchley DR, Norman JC (2002) Paxillin associates with poly(A)-binding protein 1 at the dense endoplasmic reticulum and the leading edge of migrating cells. J Biol Chem 277:6428–6437

    Article  PubMed  CAS  Google Scholar 

  • Woods AJ, Kantidakis T, Sabe H, Critchley DR, Norman JC (2005) Interaction of paxillin with poly(A)-binding protein 1 and its role in focal adhesion turnover and cell migration. Mol Cell Biol 25:3763–3773

    Article  PubMed  CAS  Google Scholar 

  • Ye J, Zhao J, Hoffmann-Rohrer U, Grummt I (2008) Nuclear myosin I acts in concert with polymeric actin to drive RNA polymerase I transcription. Genes Dev 22:322–330

    Article  PubMed  CAS  Google Scholar 

  • Young KG, Kothary R (2005) Spectrin repeat proteins in the nucleus. BioEssays 27:144–152

    Article  PubMed  CAS  Google Scholar 

  • Yuan Y, Shen Z (2001) Interaction with BRCA2 suggests a role for filamin-1 (hsFLNa) in DNA damage response. J Biol Chem 276:48318–48324

    PubMed  CAS  Google Scholar 

  • Yuan BZ, Jefferson AM, Millecchia L, Popescu NC, Reynolds SH (2007) Morphological changes and nuclear translocation of DLC1 tumor suppressor protein precede apoptosis in human non-small cell lung carcinoma cells. Exp Cell Res 313:3868–3880

    Article  PubMed  CAS  Google Scholar 

  • Zigmond SH (2004) Beginning and ending an actin filament: control at the barbed end. Curr Top Dev Biol 63:145–188

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Prof. T. Karaoli for anti-p190RhoGap antibody, Prof. M. Way for anti-paxillin antibody, Dr. Y. Raymond for anti-lamin A antibody, and Dr. Pavel Dráber for anti-tubulin antibody. We are grateful to K. Gaplovská-Kyselá for her help with writing the manuscript. This work was supported by The Ministry of Education, Youth and Sports of the Czech Republic (Ref. No. LC 545 and 2B06063), by the Grant Agency of the Czech Republic (204/05H023), and by the institutional grant AV0Z50520514.

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Dingová, H., Fukalová, J., Maninová, M. et al. Ultrastructural localization of actin and actin-binding proteins in the nucleus. Histochem Cell Biol 131, 425–434 (2009). https://doi.org/10.1007/s00418-008-0539-z

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