Chaperone activity of alpha-crystallins modulates intermediate filament assembly.
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Journal: 1994/March - EMBO Journal
ISSN: 0261-4189
PUBMED: 7906647
Abstract:
Intermediate filaments are generally regarded as one of the most insoluble and resilient cytoskeletal structures of eukaryotic cells. In extracts from the ocular lens, we noticed an unusually high level of vimentin in a soluble, non-filamentous form. Immunoprecipitation of this soluble vimentin resulted in the co-precipitation of alpha-crystallins. The alpha-crystallins are homologous to the small heat shock proteins (sHSPs) and have recently been identified as molecular chaperones, capable of preventing the heat-induced aggregation of proteins. We find that the alpha-crystallins dramatically inhibit the in vitro assembly of GFAP and vimentin in an ATP-independent manner. This inhibition is also independent of the phosphorylation state of the alpha-crystallin polypeptides and each one of the four polypeptides, either alpha A1-, alpha A2-, alpha B1- or alpha B2-crystallin, are equally effective in this inhibition. Furthermore, we show that alpha-crystallins can increase the soluble pool of GFAP when added to preformed filaments. Electron microscopy demonstrated that alpha-crystallin particles could bind to intermediate filaments in a regular fashion, the spacing coinciding with the molecular length of GFAP. This is the first report, as far as we are aware, of a chaperone being involved in intermediate filament assembly and implicates chaperones in the remodeling of intermediate filaments during development and cell differentiation.
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EMBO J 13(4): 945-953
PMID: 7906647

Chaperone activity of alpha-crystallins modulates intermediate filament assembly.

Abstract

Intermediate filaments are generally regarded as one of the most insoluble and resilient cytoskeletal structures of eukaryotic cells. In extracts from the ocular lens, we noticed an unusually high level of vimentin in a soluble, non-filamentous form. Immunoprecipitation of this soluble vimentin resulted in the co-precipitation of alpha-crystallins. The alpha-crystallins are homologous to the small heat shock proteins (sHSPs) and have recently been identified as molecular chaperones, capable of preventing the heat-induced aggregation of proteins. We find that the alpha-crystallins dramatically inhibit the in vitro assembly of GFAP and vimentin in an ATP-independent manner. This inhibition is also independent of the phosphorylation state of the alpha-crystallin polypeptides and each one of the four polypeptides, either alpha A1-, alpha A2-, alpha B1- or alpha B2-crystallin, are equally effective in this inhibition. Furthermore, we show that alpha-crystallins can increase the soluble pool of GFAP when added to preformed filaments. Electron microscopy demonstrated that alpha-crystallin particles could bind to intermediate filaments in a regular fashion, the spacing coinciding with the molecular length of GFAP. This is the first report, as far as we are aware, of a chaperone being involved in intermediate filament assembly and implicates chaperones in the remodeling of intermediate filaments during development and cell differentiation.

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Selected References

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  • Atomi Y, Yamada S, Strohman R, Nonomura Y. Alpha B-crystallin in skeletal muscle: purification and localization. J Biochem. 1991 Nov;110(5):812–822. [PubMed] [Google Scholar]
  • Bennardini F, Wrzosek A, Chiesi M. Alpha B-crystallin in cardiac tissue. Association with actin and desmin filaments. Circ Res. 1992 Aug;71(2):288–294. [PubMed] [Google Scholar]
  • Bergenhem N, Carlsson U. Denaturation and renaturation of bovine and human cobalt-carbonic anhydrases in guanidine-HCI. Ann N Y Acad Sci. 1984;429:137–139. [PubMed] [Google Scholar]
  • Bhat SP, Nagineni CN. alpha B subunit of lens-specific protein alpha-crystallin is present in other ocular and non-ocular tissues. Biochem Biophys Res Commun. 1989 Jan 16;158(1):319–325. [PubMed] [Google Scholar]
  • Bhat SP, Horwitz J, Srinivasan A, Ding L. Alpha B-crystallin exists as an independent protein in the heart and in the lens. Eur J Biochem. 1991 Dec 18;202(3):775–781. [PubMed] [Google Scholar]
  • Bloemendal H. Proctor lecture. Disorganization of membranes and abnormal intermediate filament assembly lead to cataract. Invest Ophthalmol Vis Sci. 1991 Mar;32(3):445–455. [PubMed] [Google Scholar]
  • Collier NC, Schlesinger MJ. The dynamic state of heat shock proteins in chicken embryo fibroblasts. J Cell Biol. 1986 Oct;103(4):1495–1507.[PMC free article] [PubMed] [Google Scholar]
  • Coulombe PA. The cellular and molecular biology of keratins: beginning a new era. Curr Opin Cell Biol. 1993 Feb;5(1):17–29. [PubMed] [Google Scholar]
  • Creighton TE. Effects of urea and guanidine-HCl on the folding and unfolding of pancreatic trypsin inhibitor. J Mol Biol. 1977 Jun 25;113(2):313–328. [PubMed] [Google Scholar]
  • de Jong WW, Zweers A, Versteeg M, Nuy-Terwindt EC. Primary structures of the alpha-crystallin A chains of twenty-eight mammalian species, chicken and frog. Eur J Biochem. 1984 May 15;141(1):131–140. [PubMed] [Google Scholar]
  • de Jong WW, Leunissen JA, Voorter CE. Evolution of the alpha-crystallin/small heat-shock protein family. Mol Biol Evol. 1993 Jan;10(1):103–126. [PubMed] [Google Scholar]
  • Dubin RA, Ally AH, Chung S, Piatigorsky J. Human alpha B-crystallin gene and preferential promoter function in lens. Genomics. 1990 Aug;7(4):594–601. [PubMed] [Google Scholar]
  • Ellis M, Alousi S, Lawniczak J, Maisel H, Welsh M. Studies on lens vimentin. Exp Eye Res. 1984 Feb;38(2):195–202. [PubMed] [Google Scholar]
  • Eriksson JE, Brautigan DL, Vallee R, Olmsted J, Fujiki H, Goldman RD. Cytoskeletal integrity in interphase cells requires protein phosphatase activity. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):11093–11097.[PMC free article] [PubMed] [Google Scholar]
  • FitzGerald PG, Gottlieb W. The Mr 115 kd fiber cell-specific protein is a component of the lens cytoskeleton. Curr Eye Res. 1989 Aug;8(8):801–811. [PubMed] [Google Scholar]
  • FitzGerald PG, Graham D. Ultrastructural localization of alpha A-crystallin to the bovine lens fiber cell cytoskeleton. Curr Eye Res. 1991 May;10(5):417–436. [PubMed] [Google Scholar]
  • Flaherty KM, McKay DB, Kabsch W, Holmes KC. Similarity of the three-dimensional structures of actin and the ATPase fragment of a 70-kDa heat shock cognate protein. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):5041–5045.[PMC free article] [PubMed] [Google Scholar]
  • Gao Y, Thomas JO, Chow RL, Lee GH, Cowan NJ. A cytoplasmic chaperonin that catalyzes beta-actin folding. Cell. 1992 Jun 12;69(6):1043–1050. [PubMed] [Google Scholar]
  • Geisler N, Weber K. Isolation of polymerization-competent vimentin from porcine eye lens tissue. FEBS Lett. 1981 Mar 23;125(2):253–256. [PubMed] [Google Scholar]
  • Gething MJ, Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. [PubMed] [Google Scholar]
  • Gounari F, Merdes A, Quinlan R, Hess J, FitzGerald PG, Ouzounis CA, Georgatos SD. Bovine filensin possesses primary and secondary structure similarity to intermediate filament proteins. J Cell Biol. 1993 May;121(4):847–853.[PMC free article] [PubMed] [Google Scholar]
  • Green LA, Liem RK. Beta-internexin is a microtubule-associated protein identical to the 70-kDa heat-shock cognate protein and the clathrin uncoating ATPase. J Biol Chem. 1989 Sep 15;264(26):15210–15215. [PubMed] [Google Scholar]
  • Hess JF, Casselman JT, FitzGerald PG. cDNA analysis of the 49 kDa lens fiber cell cytoskeletal protein: a new, lens-specific member of the intermediate filament family? Curr Eye Res. 1993 Jan;12(1):77–88. [PubMed] [Google Scholar]
  • Horwitz J. Alpha-crystallin can function as a molecular chaperone. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10449–10453.[PMC free article] [PubMed] [Google Scholar]
  • Inagaki M, Gonda Y, Nishizawa K, Kitamura S, Sato C, Ando S, Tanabe K, Kikuchi K, Tsuiki S, Nishi Y. Phosphorylation sites linked to glial filament disassembly in vitro locate in a non-alpha-helical head domain. J Biol Chem. 1990 Mar 15;265(8):4722–4729. [PubMed] [Google Scholar]
  • Isaacs WB, Cook RK, Van Atta JC, Redmond CM, Fulton AB. Assembly of vimentin in cultured cells varies with cell type. J Biol Chem. 1989 Oct 25;264(30):17953–17960. [PubMed] [Google Scholar]
  • Iwaki T, Kume-Iwaki A, Liem RK, Goldman JE. Alpha B-crystallin is expressed in non-lenticular tissues and accumulates in Alexander's disease brain. Cell. 1989 Apr 7;57(1):71–78. [PubMed] [Google Scholar]
  • Iwaki T, Kume-Iwaki A, Goldman JE. Cellular distribution of alpha B-crystallin in non-lenticular tissues. J Histochem Cytochem. 1990 Jan;38(1):31–39. [PubMed] [Google Scholar]
  • Kartasova T, Roop DR, Holbrook KA, Yuspa SH. Mouse differentiation-specific keratins 1 and 10 require a preexisting keratin scaffold to form a filament network. J Cell Biol. 1993 Mar;120(5):1251–1261.[PMC free article] [PubMed] [Google Scholar]
  • Kasper M, Viebahn C. Cytokeratin expression and early lens development. Anat Embryol (Berl) 1992 Aug;186(3):285–290. [PubMed] [Google Scholar]
  • Kato K, Shinohara H, Kurobe N, Goto S, Inaguma Y, Ohshima K. Immunoreactive alpha A crystallin in rat non-lenticular tissues detected with a sensitive immunoassay method. Biochim Biophys Acta. 1991 Oct 25;1080(2):173–180. [PubMed] [Google Scholar]
  • Kato K, Shinohara H, Kurobe N, Inaguma Y, Shimizu K, Ohshima K. Tissue distribution and developmental profiles of immunoreactive alpha B crystallin in the rat determined with a sensitive immunoassay system. Biochim Biophys Acta. 1991 May 24;1074(1):201–208. [PubMed] [Google Scholar]
  • Kato K, Shinohara H, Goto S, Inaguma Y, Morishita R, Asano T. Copurification of small heat shock protein with alpha B crystallin from human skeletal muscle. J Biol Chem. 1992 Apr 15;267(11):7718–7725. [PubMed] [Google Scholar]
  • Kato K, Goto S, Hasegawa K, Shinohara H, Inaguma Y. Responses to heat shock of alpha B crystallin and HSP28 in U373 MG human glioma cells. Biochim Biophys Acta. 1993 Feb 17;1175(3):257–262. [PubMed] [Google Scholar]
  • Kato S, Hirano A, Umahara T, Llena JF, Herz F, Ohama E. Ultrastructural and immunohistochemical studies on ballooned cortical neurons in Creutzfeldt-Jakob disease: expression of alpha B-crystallin, ubiquitin and stress-response protein 27. Acta Neuropathol. 1992;84(4):443–448. [PubMed] [Google Scholar]
  • Klemenz R, Fröhli E, Steiger RH, Schäfer R, Aoyama A. Alpha B-crystallin is a small heat shock protein. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3652–3656.[PMC free article] [PubMed] [Google Scholar]
  • Klemenz R, Andres AC, Fröhli E, Schäfer R, Aoyama A. Expression of the murine small heat shock proteins hsp 25 and alpha B crystallin in the absence of stress. J Cell Biol. 1993 Feb;120(3):639–645.[PMC free article] [PubMed] [Google Scholar]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed] [Google Scholar]
  • Liem RK. Molecular biology of neuronal intermediate filaments. Curr Opin Cell Biol. 1993 Feb;5(1):12–16. [PubMed] [Google Scholar]
  • Longoni S, James P, Chiesi M. Cardiac alpha-crystallin. I. Isolation and identification. Mol Cell Biochem. 1990 Dec 3;99(1):113–120. [PubMed] [Google Scholar]
  • Lowe J, McDermott H, Pike I, Spendlove I, Landon M, Mayer RJ. alpha B crystallin expression in non-lenticular tissues and selective presence in ubiquitinated inclusion bodies in human disease. J Pathol. 1992 Jan;166(1):61–68. [PubMed] [Google Scholar]
  • Merck KB, Groenen PJ, Voorter CE, de Haard-Hoekman WA, Horwitz J, Bloemendal H, de Jong WW. Structural and functional similarities of bovine alpha-crystallin and mouse small heat-shock protein. A family of chaperones. J Biol Chem. 1993 Jan 15;268(2):1046–1052. [PubMed] [Google Scholar]
  • Miron T, Vancompernolle K, Vandekerckhove J, Wilchek M, Geiger B. A 25-kD inhibitor of actin polymerization is a low molecular mass heat shock protein. J Cell Biol. 1991 Jul;114(2):255–261.[PMC free article] [PubMed] [Google Scholar]
  • Osborn M, Debus E, Weber K. Monoclonal antibodies specific for vimentin. Eur J Cell Biol. 1984 May;34(1):137–143. [PubMed] [Google Scholar]
  • Osborn M, Altmannsberger M, Debus E, Weber K. Differentiation of the major human tumor groups using conventional and monoclonal antibodies specific for individual intermediate filament proteins. Ann N Y Acad Sci. 1985;455:649–668. [PubMed] [Google Scholar]
  • Quinlan RA, Schiller DL, Hatzfeld M, Achtstätter T, Moll R, Jorcano JL, Magin TM, Franke WW. Patterns of expression and organization of cytokeratin intermediate filaments. Ann N Y Acad Sci. 1985;455:282–306. [PubMed] [Google Scholar]
  • Quinlan RA, Moir RD, Stewart M. Expression in Escherichia coli of fragments of glial fibrillary acidic protein: characterization, assembly properties and paracrystal formation. J Cell Sci. 1989 May;93(Pt 1):71–83. [PubMed] [Google Scholar]
  • Quinlan RA, Carter JM, Hutcheson AM, Campbell DG. The 53kDa polypeptide component of the bovine fibre cell cytoskeleton is derived from the 115kDa beaded filament protein: evidence for a fibre cell specific intermediate filament protein. Curr Eye Res. 1992 Sep;11(9):909–921. [PubMed] [Google Scholar]
  • Ramaekers FC, Dunia I, Dodemont HJ, Benedetti EL, Bloemendal H. Lenticular intermediate-sized filaments: biosynthesis and interaction with plasma membrane. Proc Natl Acad Sci U S A. 1982 May;79(10):3208–3212.[PMC free article] [PubMed] [Google Scholar]
  • Remington SG. Chicken filensin: a lens fiber cell protein that exhibits sequence similarity to intermediate filament proteins. J Cell Sci. 1993 Aug;105(Pt 4):1057–1068. [PubMed] [Google Scholar]
  • Scotting P, McDermott H, Mayer RJ. Ubiquitin-protein conjugates and alpha B crystallin are selectively present in cells undergoing major cytomorphological reorganisation in early chicken embryos. FEBS Lett. 1991 Jul 8;285(1):75–79. [PubMed] [Google Scholar]
  • Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillessen D, Gabbiani G. A monoclonal antibody against alpha-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol. 1986 Dec;103(6 Pt 2):2787–2796.[PMC free article] [PubMed] [Google Scholar]
  • Soellner P, Quinlan RA, Franke WW. Identification of a distinct soluble subunit of an intermediate filament protein: tetrameric vimentin from living cells. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7929–7933.[PMC free article] [PubMed] [Google Scholar]
  • Srinivasan AN, Nagineni CN, Bhat SP. alpha A-crystallin is expressed in non-ocular tissues. J Biol Chem. 1992 Nov 15;267(32):23337–23341. [PubMed] [Google Scholar]
  • Stewart M. Intermediate filament structure and assembly. Curr Opin Cell Biol. 1993 Feb;5(1):3–11. [PubMed] [Google Scholar]
  • Voorter CE, Wintjes L, Bloemendal H, de Jong WW. Relocalization of alpha B-crystallin by heat shock in ovarian carcinoma cells. FEBS Lett. 1992 Sep 7;309(2):111–114. [PubMed] [Google Scholar]
  • Waseem NH, Lane DP. Monoclonal antibody analysis of the proliferating cell nuclear antigen (PCNA). Structural conservation and the detection of a nucleolar form. J Cell Sci. 1990 May;96(Pt 1):121–129. [PubMed] [Google Scholar]
  • Welch WJ, Feramisco JR. Disruption of the three cytoskeletal networks in mammalian cells does not affect transcription, translation, or protein translocation changes induced by heat shock. Mol Cell Biol. 1985 Jul;5(7):1571–1581.[PMC free article] [PubMed] [Google Scholar]
  • Welch WJ, Suhan JP. Morphological study of the mammalian stress response: characterization of changes in cytoplasmic organelles, cytoskeleton, and nucleoli, and appearance of intranuclear actin filaments in rat fibroblasts after heat-shock treatment. J Cell Biol. 1985 Oct;101(4):1198–1211.[PMC free article] [PubMed] [Google Scholar]
  • Wessel D, Flügge UI. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984 Apr;138(1):141–143. [PubMed] [Google Scholar]
  • Wiech H, Buchner J, Zimmermann R, Jakob U. Hsp90 chaperones protein folding in vitro. Nature. 1992 Jul 9;358(6382):169–170. [PubMed] [Google Scholar]
  • Yaffe MB, Farr GW, Miklos D, Horwich AL, Sternlicht ML, Sternlicht H. TCP1 complex is a molecular chaperone in tubulin biogenesis. Nature. 1992 Jul 16;358(6383):245–248. [PubMed] [Google Scholar]
Department of Biochemistry, The University, Dundee, UK.
Department of Biochemistry, The University, Dundee, UK.
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Abstract
Intermediate filaments are generally regarded as one of the most insoluble and resilient cytoskeletal structures of eukaryotic cells. In extracts from the ocular lens, we noticed an unusually high level of vimentin in a soluble, non-filamentous form. Immunoprecipitation of this soluble vimentin resulted in the co-precipitation of alpha-crystallins. The alpha-crystallins are homologous to the small heat shock proteins (sHSPs) and have recently been identified as molecular chaperones, capable of preventing the heat-induced aggregation of proteins. We find that the alpha-crystallins dramatically inhibit the in vitro assembly of GFAP and vimentin in an ATP-independent manner. This inhibition is also independent of the phosphorylation state of the alpha-crystallin polypeptides and each one of the four polypeptides, either alpha A1-, alpha A2-, alpha B1- or alpha B2-crystallin, are equally effective in this inhibition. Furthermore, we show that alpha-crystallins can increase the soluble pool of GFAP when added to preformed filaments. Electron microscopy demonstrated that alpha-crystallin particles could bind to intermediate filaments in a regular fashion, the spacing coinciding with the molecular length of GFAP. This is the first report, as far as we are aware, of a chaperone being involved in intermediate filament assembly and implicates chaperones in the remodeling of intermediate filaments during development and cell differentiation.
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