• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Oct 1, 2001; 359(Pt 1): 55–64.
PMCID: PMC1222121

Chloride intracellular channel protein CLIC4 (p64H1) binds directly to brain dynamin I in a complex containing actin, tubulin and 14-3-3 isoforms.

Abstract

Mammalian chloride intracellular channel (CLIC) (p64-related) proteins are widely expressed, with an unusual dual localization as both soluble and integral membrane proteins. The molecular basis for their cellular localization and ion channel activity remains unclear. To help in addressing these problems, we identified novel rat brain CLIC4 (p64H1) binding partners by affinity chromatography, mass spectrometric analysis and microsequencing. Brain CLIC4 binds dynamin I, alpha-tubulin, beta-actin, creatine kinase and two 14-3-3 isoforms; the interactions are confirmed in vivo by immunoprecipitation. Gel overlay and reverse pull-down assays indicate that the binding of CLIC4 to dynamin I and 14-3-3zeta is direct. In HEK-293 cells, biochemical and immunofluorescence analyses show partial co-localization of recombinant CLIC4 with caveolin and with functional caveolae, which is consistent with a dynamin-associated role for CLIC4 in caveolar endocytosis. We speculate that brain CLIC4 might be involved in the dynamics of neuronal plasma membrane microdomains (micropatches) containing caveolin-like proteins and might also have other cellular roles related to membrane trafficking. Our results provide the basis for new hypotheses concerning novel ways in which CLIC proteins might be associated with cell membrane remodelling, the control of cell shape, and anion channel activity.

Full Text

The Full Text of this article is available as a PDF (310K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Michikawa T, Miyawaki A, Furuichi T, Mikoshiba K. Inositol 1,4,5-trisphosphate receptors and calcium signaling. Crit Rev Neurobiol. 1996;10(1):39–55. [PubMed]
  • Shoshan-Barmatz V, Ashley RH. The structure, function, and cellular regulation of ryanodine-sensitive Ca2+ release channels. Int Rev Cytol. 1998;183:185–270. [PubMed]
  • Simon SM, Blobel G. A protein-conducting channel in the endoplasmic reticulum. Cell. 1991 May 3;65(3):371–380. [PubMed]
  • Tabares L, Mazzanti M, Clapham DE. Chloride channels in the nuclear membrane. J Membr Biol. 1991 Jul;123(1):49–54. [PubMed]
  • Ballarin C, Sorgato MC. Anion channels of the inner membrane of mammalian and yeast mitochondria. J Bioenerg Biomembr. 1996 Apr;28(2):125–130. [PubMed]
  • Glickman J, Croen K, Kelly S, Al-Awqati Q. Golgi membranes contain an electrogenic H+ pump in parallel to a chloride conductance. J Cell Biol. 1983 Oct;97(4):1303–1308. [PMC free article] [PubMed]
  • Nordeen MH, Jones SM, Howell KE, Caldwell JH. GOLAC: an endogenous anion channel of the Golgi complex. Biophys J. 2000 Jun;78(6):2918–2928. [PMC free article] [PubMed]
  • Stanley EF, Ehrenstein G, Russell JT. Evidence for anion channels in secretory vesicles. Neuroscience. 1988 Jun;25(3):1035–1039. [PubMed]
  • Arispe N, Pollard HB, Rojas E. Calcium-independent K(+)-selective channel from chromaffin granule membranes. J Membr Biol. 1992 Nov;130(2):191–202. [PubMed]
  • Ashley RH, Brown DM, Apps DK, Phillips JH. Evidence for a K+ channel in bovine chromaffin granule membranes: single-channel properties and possible bioenergetic significance. Eur Biophys J. 1994;23(4):263–275. [PubMed]
  • al-Awqati Q. Chloride channels of intracellular organelles. Curr Opin Cell Biol. 1995 Aug;7(4):504–508. [PubMed]
  • Nilius B, Eggermont J, Voets T, Droogmans G. Volume-activated Cl- channels. Gen Pharmacol. 1996 Oct;27(7):1131–1140. [PubMed]
  • Okada Y. Volume expansion-sensing outward-rectifier Cl- channel: fresh start to the molecular identity and volume sensor. Am J Physiol. 1997 Sep;273(3 Pt 1):C755–C789. [PubMed]
  • Landry D, Sullivan S, Nicolaides M, Redhead C, Edelman A, Field M, al-Awqati Q, Edwards J. Molecular cloning and characterization of p64, a chloride channel protein from kidney microsomes. J Biol Chem. 1993 Jul 15;268(20):14948–14955. [PubMed]
  • Howell S, Duncan RR, Ashley RH. Identification and characterisation of a homologue of p64 in rat tissues. FEBS Lett. 1996 Jul 22;390(2):207–210. [PubMed]
  • Duncan RR, Westwood PK, Boyd A, Ashley RH. Rat brain p64H1, expression of a new member of the p64 chloride channel protein family in endoplasmic reticulum. J Biol Chem. 1997 Sep 19;272(38):23880–23886. [PubMed]
  • Redhead C, Sullivan SK, Koseki C, Fujiwara K, Edwards JC. Subcellular distribution and targeting of the intracellular chloride channel p64. Mol Biol Cell. 1997 Apr;8(4):691–704. [PMC free article] [PubMed]
  • Valenzuela SM, Martin DK, Por SB, Robbins JM, Warton K, Bootcov MR, Schofield PR, Campbell TJ, Breit SN. Molecular cloning and expression of a chloride ion channel of cell nuclei. J Biol Chem. 1997 May 9;272(19):12575–12582. [PubMed]
  • Qian Z, Okuhara D, Abe MK, Rosner MR. Molecular cloning and characterization of a mitogen-activated protein kinase-associated intracellular chloride channel. J Biol Chem. 1999 Jan 15;274(3):1621–1627. [PubMed]
  • Chuang JZ, Milner TA, Zhu M, Sung CH. A 29 kDa intracellular chloride channel p64H1 is associated with large dense-core vesicles in rat hippocampal neurons. J Neurosci. 1999 Apr 15;19(8):2919–2928. [PubMed]
  • Edwards JC. A novel p64-related Cl- channel: subcellular distribution and nephron segment-specific expression. Am J Physiol. 1999 Mar;276(3 Pt 2):F398–F408. [PubMed]
  • Fernández-Salas E, Sagar M, Cheng C, Yuspa SH, Weinberg WC. p53 and tumor necrosis factor alpha regulate the expression of a mitochondrial chloride channel protein. J Biol Chem. 1999 Dec 17;274(51):36488–36497. [PubMed]
  • Nishizawa T, Nagao T, Iwatsubo T, Forte JG, Urushidani T. Molecular cloning and characterization of a novel chloride intracellular channel-related protein, parchorin, expressed in water-secreting cells. J Biol Chem. 2000 Apr 14;275(15):11164–11173. [PubMed]
  • Heiss NS, Poustka A. Genomic structure of a novel chloride channel gene, CLIC2, in Xq28. Genomics. 1997 Oct 1;45(1):224–228. [PubMed]
  • Berryman M, Bretscher A. Identification of a novel member of the chloride intracellular channel gene family (CLIC5) that associates with the actin cytoskeleton of placental microvilli. Mol Biol Cell. 2000 May;11(5):1509–1521. [PMC free article] [PubMed]
  • Tulk BM, Schlesinger PH, Kapadia SA, Edwards JC. CLIC-1 functions as a chloride channel when expressed and purified from bacteria. J Biol Chem. 2000 Sep 1;275(35):26986–26993. [PubMed]
  • Strange K, Emma F, Jackson PS. Cellular and molecular physiology of volume-sensitive anion channels. Am J Physiol. 1996 Mar;270(3 Pt 1):C711–C730. [PubMed]
  • Witke W, Podtelejnikov AV, Di Nardo A, Sutherland JD, Gurniak CB, Dotti C, Mann M. In mouse brain profilin I and profilin II associate with regulators of the endocytic pathway and actin assembly. EMBO J. 1998 Feb 16;17(4):967–976. [PMC free article] [PubMed]
  • Spudich JA, Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed]
  • van der Bliek AM. Functional diversity in the dynamin family. Trends Cell Biol. 1999 Mar;9(3):96–102. [PubMed]
  • McNiven MA, Cao H, Pitts KR, Yoon Y. The dynamin family of mechanoenzymes: pinching in new places. Trends Biochem Sci. 2000 Mar;25(3):115–120. [PubMed]
  • Oh P, McIntosh DP, Schnitzer JE. Dynamin at the neck of caveolae mediates their budding to form transport vesicles by GTP-driven fission from the plasma membrane of endothelium. J Cell Biol. 1998 Apr 6;141(1):101–114. [PMC free article] [PubMed]
  • Henley JR, Krueger EW, Oswald BJ, McNiven MA. Dynamin-mediated internalization of caveolae. J Cell Biol. 1998 Apr 6;141(1):85–99. [PMC free article] [PubMed]
  • Pike LJ, Casey L. Localization and turnover of phosphatidylinositol 4,5-bisphosphate in caveolin-enriched membrane domains. J Biol Chem. 1996 Oct 25;271(43):26453–26456. [PubMed]
  • Redhead CR, Edelman AE, Brown D, Landry DW, al-Awqati Q. A ubiquitous 64-kDa protein is a component of a chloride channel of plasma and intracellular membranes. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3716–3720. [PMC free article] [PubMed]
  • Landry DW, Akabas MH, Redhead C, Edelman A, Cragoe EJ, Jr, Al-Awqati Q. Purification and reconstitution of chloride channels from kidney and trachea. Science. 1989 Jun 23;244(4911):1469–1472. [PubMed]
  • Edwards JC, Tulk B, Schlesinger PH. Functional expression of p64, an intracellular chloride channel protein. J Membr Biol. 1998 May 15;163(2):119–127. [PubMed]
  • Obar RA, Collins CA, Hammarback JA, Shpetner HS, Vallee RB. Molecular cloning of the microtubule-associated mechanochemical enzyme dynamin reveals homology with a new family of GTP-binding proteins. Nature. 1990 Sep 20;347(6290):256–261. [PubMed]
  • Schmid SL, McNiven MA, De Camilli P. Dynamin and its partners: a progress report. Curr Opin Cell Biol. 1998 Aug;10(4):504–512. [PubMed]
  • Sotgia F, Lee JK, Das K, Bedford M, Petrucci TC, Macioce P, Sargiacomo M, Bricarelli FD, Minetti C, Sudol M, et al. Caveolin-3 directly interacts with the C-terminal tail of beta -dystroglycan. Identification of a central WW-like domain within caveolin family members. J Biol Chem. 2000 Dec 1;275(48):38048–38058. [PubMed]
  • Tettamanti G, Riboni L. Gangliosides and modulation of the function of neural cells. Adv Lipid Res. 1993;25:235–267. [PubMed]
  • Lang DM, Lommel S, Jung M, Ankerhold R, Petrausch B, Laessing U, Wiechers MF, Plattner H, Stuermer CA. Identification of reggie-1 and reggie-2 as plasmamembrane-associated proteins which cocluster with activated GPI-anchored cell adhesion molecules in non-caveolar micropatches in neurons. J Neurobiol. 1998 Dec;37(4):502–523. [PubMed]
  • Cook TA, Urrutia R, McNiven MA. Identification of dynamin 2, an isoform ubiquitously expressed in rat tissues. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):644–648. [PMC free article] [PubMed]
  • Torre E, McNiven MA, Urrutia R. Dynamin 1 antisense oligonucleotide treatment prevents neurite formation in cultured hippocampal neurons. J Biol Chem. 1994 Dec 23;269(51):32411–32417. [PubMed]
  • Fish KN, Schmid SL, Damke H. Evidence that dynamin-2 functions as a signal-transducing GTPase. J Cell Biol. 2000 Jul 10;150(1):145–154. [PMC free article] [PubMed]
  • Palestini P, Pitto M, Tedeschi G, Ferraretto A, Parenti M, Brunner J, Masserini M. Tubulin anchoring to glycolipid-enriched, detergent-resistant domains of the neuronal plasma membrane. J Biol Chem. 2000 Apr 7;275(14):9978–9985. [PubMed]
  • Aitken A. 14-3-3 and its possible role in co-ordinating multiple signalling pathways. Trends Cell Biol. 1996 Sep;6(9):341–347. [PubMed]
  • Martin H, Rostas J, Patel Y, Aitken A. Subcellular localisation of 14-3-3 isoforms in rat brain using specific antibodies. J Neurochem. 1994 Dec;63(6):2259–2265. [PubMed]
  • Lang T, Wacker I, Wunderlich I, Rohrbach A, Giese G, Soldati T, Almers W. Role of actin cortex in the subplasmalemmal transport of secretory granules in PC-12 cells. Biophys J. 2000 Jun;78(6):2863–2877. [PMC free article] [PubMed]
  • Lamaze C, Fujimoto LM, Yin HL, Schmid SL. The actin cytoskeleton is required for receptor-mediated endocytosis in mammalian cells. J Biol Chem. 1997 Aug 15;272(33):20332–20335. [PubMed]
  • McNiven MA, Kim L, Krueger EW, Orth JD, Cao H, Wong TW. Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. J Cell Biol. 2000 Oct 2;151(1):187–198. [PMC free article] [PubMed]
  • Trouet D, Nilius B, Jacobs A, Remacle C, Droogmans G, Eggermont J. Caveolin-1 modulates the activity of the volume-regulated chloride channel. J Physiol. 1999 Oct 1;520(Pt 1):113–119. [PMC free article] [PubMed]
  • Parton RG, Joggerst B, Simons K. Regulated internalization of caveolae. J Cell Biol. 1994 Dec;127(5):1199–1215. [PMC free article] [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Gene
    Gene
    Gene links
  • Gene (nucleotide)
    Gene (nucleotide)
    Records in Gene identified from shared sequence links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • HomoloGene
    HomoloGene
    HomoloGene links
  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...