Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 Aug 2;114(4):773–786. doi: 10.1083/jcb.114.4.773

A potential role for the COOH-terminal domain in the lateral packing of type III intermediate filaments

PMCID: PMC2289903  PMID: 1714461

Abstract

To identify sites of self-association in type III intermediate filament (IF) proteins, we have taken an "anti-idiotypic antibody" approach. A mAb (anti-Ct), recognizing a similar feature near the end of the rod domain of vimentin, desmin, and peripherin (epsilon site or epsilon epitope), was characterized. Anti-idiotypic antibodies, generated by immunizing rabbits with purified anti-Ct, recognize a site (presumably "complementary" to the epsilon epitope) common among vimentin, desmin, and peripherin (beta site or beta epitope). The beta epitope is represented in a synthetic peptide (PII) modeled after the 30 COOH- terminal residues of peripherin, as seen by comparative immunoblotting assays. Consistent with the idea of an association between the epsilon and the beta site, PII binds in vitro to intact IF proteins and fragments containing the epsilon epitope, but not to IF proteins that do not react with anti-Ct. Microinjection experiments conducted in vivo and filament reconstitution assays carried out in vitro further demonstrate that "uncoupling" of this site-specific association (by competition with PII or anti-Ct) interferes with normal IF architecture, resulting in the formation of filaments and filament bundles with diameters much greater than that of the normal IFs. These thick fibers are very similar to the ones observed previously when a derivative of desmin missing 27 COOH-terminal residues was assembled in vitro (Kaufmann, E., K. Weber, and N. Geisler. 1985. J. Mol. Biol. 185:733-742). As a molecular explanation, we propose here that the epsilon and the beta sites of type III IF proteins are "complementary" and associate during filament assembly. As a result of this association, we further postulate the formation of a surface-exposed "loop" or "hairpin" structure that may sterically prevent inappropriate filament-filament aggregation and regulate filament thickness.

Full Text

The Full Text of this article is available as a PDF (3.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aebi U., Cohn J., Buhle L., Gerace L. The nuclear lamina is a meshwork of intermediate-type filaments. Nature. 1986 Oct 9;323(6088):560–564. doi: 10.1038/323560a0. [DOI] [PubMed] [Google Scholar]
  2. Albers K., Fuchs E. Expression of mutant keratin cDNAs in epithelial cells reveals possible mechanisms for initiation and assembly of intermediate filaments. J Cell Biol. 1989 Apr;108(4):1477–1493. doi: 10.1083/jcb.108.4.1477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Albers K., Fuchs E. The expression of mutant epidermal keratin cDNAs transfected in simple epithelial and squamous cell carcinoma lines. J Cell Biol. 1987 Aug;105(2):791–806. doi: 10.1083/jcb.105.2.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ansorge W., Pepperkok R. Performance of an automated system for capillary microinjection into living cells. J Biochem Biophys Methods. 1988 Aug;16(4):283–292. doi: 10.1016/0165-022x(88)90062-0. [DOI] [PubMed] [Google Scholar]
  5. Birkenberger L., Ip W. Properties of the desmin tail domain: studies using synthetic peptides and antipeptide antibodies. J Cell Biol. 1990 Nov;111(5 Pt 1):2063–2075. doi: 10.1083/jcb.111.5.2063. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coulombe P. A., Chan Y. M., Albers K., Fuchs E. Deletions in epidermal keratins leading to alterations in filament organization in vivo and in intermediate filament assembly in vitro. J Cell Biol. 1990 Dec;111(6 Pt 2):3049–3064. doi: 10.1083/jcb.111.6.3049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coulombe P. A., Fuchs E. Elucidating the early stages of keratin filament assembly. J Cell Biol. 1990 Jul;111(1):153–169. doi: 10.1083/jcb.111.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dale B. A., Holbrook K. A., Steinert P. M. Assembly of stratum corneum basic protein and keratin filaments in macrofibrils. Nature. 1978 Dec 14;276(5689):729–731. doi: 10.1038/276729a0. [DOI] [PubMed] [Google Scholar]
  9. Danscher G. Histochemical demonstration of heavy metals. A revised version of the sulphide silver method suitable for both light and electronmicroscopy. Histochemistry. 1981;71(1):1–16. doi: 10.1007/BF00592566. [DOI] [PubMed] [Google Scholar]
  10. Djabali K., Portier M. M., Gros F., Blobel G., Georgatos S. D. Network antibodies identify nuclear lamin B as a physiological attachment site for peripherin intermediate filaments. Cell. 1991 Jan 11;64(1):109–121. doi: 10.1016/0092-8674(91)90213-i. [DOI] [PubMed] [Google Scholar]
  11. Engel A., Eichner R., Aebi U. Polymorphism of reconstituted human epidermal keratin filaments: determination of their mass-per-length and width by scanning transmission electron microscopy (STEM). J Ultrastruct Res. 1985 Mar;90(3):323–335. doi: 10.1016/s0022-5320(85)80010-1. [DOI] [PubMed] [Google Scholar]
  12. Geisler N., Kaufmann E., Fischer S., Plessmann U., Weber K. Neurofilament architecture combines structural principles of intermediate filaments with carboxy-terminal extensions increasing in size between triplet proteins. EMBO J. 1983;2(8):1295–1302. doi: 10.1002/j.1460-2075.1983.tb01584.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geisler N., Kaufmann E., Weber K. Antiparallel orientation of the two double-stranded coiled-coils in the tetrameric protofilament unit of intermediate filaments. J Mol Biol. 1985 Mar 5;182(1):173–177. doi: 10.1016/0022-2836(85)90035-x. [DOI] [PubMed] [Google Scholar]
  14. Geisler N., Kaufmann E., Weber K. Proteinchemical characterization of three structurally distinct domains along the protofilament unit of desmin 10 nm filaments. Cell. 1982 Aug;30(1):277–286. doi: 10.1016/0092-8674(82)90033-2. [DOI] [PubMed] [Google Scholar]
  15. Geisler N., Weber K. The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins. EMBO J. 1982;1(12):1649–1656. doi: 10.1002/j.1460-2075.1982.tb01368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Georgatos S. D., Blobel G. Lamin B constitutes an intermediate filament attachment site at the nuclear envelope. J Cell Biol. 1987 Jul;105(1):117–125. doi: 10.1083/jcb.105.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Georgatos S. D., Blobel G. Two distinct attachment sites for vimentin along the plasma membrane and the nuclear envelope in avian erythrocytes: a basis for a vectorial assembly of intermediate filaments. J Cell Biol. 1987 Jul;105(1):105–115. doi: 10.1083/jcb.105.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Georgatos S. D., Weber K., Geisler N., Blobel G. Binding of two desmin derivatives to the plasma membrane and the nuclear envelope of avian erythrocytes: evidence for a conserved site-specificity in intermediate filament-membrane interactions. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6780–6784. doi: 10.1073/pnas.84.19.6780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gill S. R., Wong P. C., Monteiro M. J., Cleveland D. W. Assembly properties of dominant and recessive mutations in the small mouse neurofilament (NF-L) subunit. J Cell Biol. 1990 Nov;111(5 Pt 1):2005–2019. doi: 10.1083/jcb.111.5.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hanukoglu I., Fuchs E. The cDNA sequence of a Type II cytoskeletal keratin reveals constant and variable structural domains among keratins. Cell. 1983 Jul;33(3):915–924. doi: 10.1016/0092-8674(83)90034-x. [DOI] [PubMed] [Google Scholar]
  21. Hatzfeld M., Weber K. Tailless keratins assemble into regular intermediate filaments in vitro. J Cell Sci. 1990 Oct;97(Pt 2):317–324. doi: 10.1242/jcs.97.2.317. [DOI] [PubMed] [Google Scholar]
  22. Hisanaga S., Hirokawa N. Structure of the peripheral domains of neurofilaments revealed by low angle rotary shadowing. J Mol Biol. 1988 Jul 20;202(2):297–305. doi: 10.1016/0022-2836(88)90459-7. [DOI] [PubMed] [Google Scholar]
  23. Höger T. H., Krohne G., Franke W. W. Amino acid sequence and molecular characterization of murine lamin B as deduced from cDNA clones. Eur J Cell Biol. 1988 Dec;47(2):283–290. [PubMed] [Google Scholar]
  24. Ip W. Modulation of desmin intermediate filament assembly by a monoclonal antibody. J Cell Biol. 1988 Mar;106(3):735–745. doi: 10.1083/jcb.106.3.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jerne N. K. The generative grammar of the immune system. EMBO J. 1985 Apr;4(4):847–852. doi: 10.1002/j.1460-2075.1985.tb03709.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jerne N. K. Towards a network theory of the immune system. Ann Immunol (Paris) 1974 Jan;125C(1-2):373–389. [PubMed] [Google Scholar]
  27. Kaufmann E., Weber K., Geisler N. Intermediate filament forming ability of desmin derivatives lacking either the amino-terminal 67 or the carboxy-terminal 27 residues. J Mol Biol. 1985 Oct 20;185(4):733–742. doi: 10.1016/0022-2836(85)90058-0. [DOI] [PubMed] [Google Scholar]
  28. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  29. Landon F., Lemonnier M., Benarous R., Huc C., Fiszman M., Gros F., Portier M. M. Multiple mRNAs encode peripherin, a neuronal intermediate filament protein. EMBO J. 1989 Jun;8(6):1719–1726. doi: 10.1002/j.1460-2075.1989.tb03564.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  31. Leonard D. G., Gorham J. D., Cole P., Greene L. A., Ziff E. B. A nerve growth factor-regulated messenger RNA encodes a new intermediate filament protein. J Cell Biol. 1988 Jan;106(1):181–193. doi: 10.1083/jcb.106.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lu X., Lane E. B. Retrovirus-mediated transgenic keratin expression in cultured fibroblasts: specific domain functions in keratin stabilization and filament formation. Cell. 1990 Aug 24;62(4):681–696. doi: 10.1016/0092-8674(90)90114-t. [DOI] [PubMed] [Google Scholar]
  33. Magin T. M., Hatzfeld M., Franke W. W. Analysis of cytokeratin domains by cloning and expression of intact and deleted polypeptides in Escherichia coli. EMBO J. 1987 Sep;6(9):2607–2615. doi: 10.1002/j.1460-2075.1987.tb02551.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Matteoni R., Kreis T. E. Translocation and clustering of endosomes and lysosomes depends on microtubules. J Cell Biol. 1987 Sep;105(3):1253–1265. doi: 10.1083/jcb.105.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nelson W. J., Traub P. Purification of the intermediate filament protein vimentin from Ehrlich ascites tumor cells. J Biol Chem. 1982 May 25;257(10):5536–5543. [PubMed] [Google Scholar]
  36. Pain D., Murakami H., Blobel G. Identification of a receptor for protein import into mitochondria. Nature. 1990 Oct 4;347(6292):444–449. doi: 10.1038/347444a0. [DOI] [PubMed] [Google Scholar]
  37. Perides G., Kühn S., Scherbarth A., Traub P. Probing of the structural stability of vimentin and desmin-type intermediate filaments with Ca2+-activated proteinase, thrombin and lysine-specific endoproteinase Lys-C. Eur J Cell Biol. 1987 Jun;43(3):450–458. [PubMed] [Google Scholar]
  38. Pruss R. M., Mirsky R., Raff M. C., Thorpe R., Dowding A. J., Anderton B. H. All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody. Cell. 1981 Dec;27(3 Pt 2):419–428. doi: 10.1016/0092-8674(81)90383-4. [DOI] [PubMed] [Google Scholar]
  39. Quax W., Egberts W. V., Hendriks W., Quax-Jeuken Y., Bloemendal H. The structure of the vimentin gene. Cell. 1983 Nov;35(1):215–223. doi: 10.1016/0092-8674(83)90224-6. [DOI] [PubMed] [Google Scholar]
  40. Quax W., van den Broek L., Egberts W. V., Ramaekers F., Bloemendal H. Characterization of the hamster desmin gene: expression and formation of desmin filaments in nonmuscle cells after gene transfer. Cell. 1985 Nov;43(1):327–338. doi: 10.1016/0092-8674(85)90038-8. [DOI] [PubMed] [Google Scholar]
  41. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Renner W., Franke W. W., Schmid E., Geisler N., Weber K., Mandelkow E. Reconstitution of intermediate-sized filaments from denatured monomeric vimentin. J Mol Biol. 1981 Jun 25;149(2):285–306. doi: 10.1016/0022-2836(81)90303-x. [DOI] [PubMed] [Google Scholar]
  43. Rivas C. I., Vera J. C., Maccioni R. B. Anti-idiotypic antibodies that react with microtubule-associated proteins are present in the sera of rabbits immunized with synthetic peptides from tubulin's regulatory domain. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6092–6096. doi: 10.1073/pnas.85.16.6092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Shechter Y., Elias D., Maron R., Cohen I. R. Mouse antibodies to the insulin receptor developing spontaneously as anti-idiotypes. I. Characterization of the antibodies. J Biol Chem. 1984 May 25;259(10):6411–6415. [PubMed] [Google Scholar]
  45. Shechter Y., Maron R., Elias D., Cohen I. R. Autoantibodies to insulin receptor spontaneously develop as anti-idiotypes in mice immunized with insulin. Science. 1982 Apr 30;216(4545):542–545. doi: 10.1126/science.7041258. [DOI] [PubMed] [Google Scholar]
  46. Shoeman R. L., Mothes E., Kesselmeier C., Traub P. Intermediate filament assembly and stability in vitro: effect and implications of the removal of head and tail domains of vimentin by the human immunodeficiency virus type 1 protease. Cell Biol Int Rep. 1990 Jul;14(7):583–594. doi: 10.1016/0309-1651(90)90038-z. [DOI] [PubMed] [Google Scholar]
  47. Steinert P. M., Rice R. H., Roop D. R., Trus B. L., Steven A. C. Complete amino acid sequence of a mouse epidermal keratin subunit and implications for the structure of intermediate filaments. Nature. 1983 Apr 28;302(5911):794–800. doi: 10.1038/302794a0. [DOI] [PubMed] [Google Scholar]
  48. Steinert P. M., Roop D. R. Molecular and cellular biology of intermediate filaments. Annu Rev Biochem. 1988;57:593–625. doi: 10.1146/annurev.bi.57.070188.003113. [DOI] [PubMed] [Google Scholar]
  49. Steven A. C., Hainfeld J. F., Trus B. L., Wall J. S., Steinert P. M. Epidermal keratin filaments assembled in vitro have masses-per-unit-length that scale according to average subunit mass: structural basis for homologous packing of subunits in intermediate filaments. J Cell Biol. 1983 Dec;97(6):1939–1944. doi: 10.1083/jcb.97.6.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Steven A. C., Hainfeld J. F., Trus B. L., Wall J. S., Steinert P. M. The distribution of mass in heteropolymer intermediate filaments assembled in vitro. Stem analysis of vimentin/desmin and bovine epidermal keratin. J Biol Chem. 1983 Jul 10;258(13):8323–8329. [PubMed] [Google Scholar]
  51. Steven A. C., Wall J., Hainfeld J., Steinert P. M. Structure of fibroblastic intermediate filaments: analysis of scanning transmission electron microscopy. Proc Natl Acad Sci U S A. 1982 May;79(10):3101–3105. doi: 10.1073/pnas.79.10.3101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Stewart M., Quinlan R. A., Moir R. D. Molecular interactions in paracrystals of a fragment corresponding to the alpha-helical coiled-coil rod portion of glial fibrillary acidic protein: evidence for an antiparallel packing of molecules and polymorphism related to intermediate filament structure. J Cell Biol. 1989 Jul;109(1):225–234. doi: 10.1083/jcb.109.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Traub P., Vorgias C. E. Involvement of the N-terminal polypeptide of vimentin in the formation of intermediate filaments. J Cell Sci. 1983 Sep;63:43–67. doi: 10.1242/jcs.63.1.43. [DOI] [PubMed] [Google Scholar]
  54. Tölle H. G., Weber K., Osborn M. Microinjection of monoclonal antibodies to vimentin, desmin, and GFA in cells which contain more than one IF type. Exp Cell Res. 1986 Feb;162(2):462–474. doi: 10.1016/0014-4827(86)90350-2. [DOI] [PubMed] [Google Scholar]
  55. Vaux D., Tooze J., Fuller S. Identification by anti-idiotype antibodies of an intracellular membrane protein that recognizes a mammalian endoplasmic reticulum retention signal. Nature. 1990 Jun 7;345(6275):495–502. doi: 10.1038/345495a0. [DOI] [PubMed] [Google Scholar]
  56. Wong P. C., Cleveland D. W. Characterization of dominant and recessive assembly-defective mutations in mouse neurofilament NF-M. J Cell Biol. 1990 Nov;111(5 Pt 1):1987–2003. doi: 10.1083/jcb.111.5.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Zackroff R. V., Goldman R. D. In vitro assembly of intermediate filaments from baby hamster kidney (BHK-21) cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6226–6230. doi: 10.1073/pnas.76.12.6226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zehner Z. E., Li Y., Roe B. A., Paterson B. M., Sax C. M. The chicken vimentin gene. Nucleotide sequence, regulatory elements, and comparison to the hamster gene. J Biol Chem. 1987 Jun 15;262(17):8112–8120. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES