Abstract
We have introduced a hybrid gene, pVVim2, composed of the 5' region of the hamster vimentin gene encoding the head and rod domain of vimentin and the 3' region of the hamster desmin gene encoding the tail domain of desmin, into the germ line of mice by pronuclear injection. RNA and protein analysis of mice transgenic for this construct showed that the pVVim2 gene was expressed at high levels in a developmental and tissue-specific manner. This indicates that the vimentin-derived segment of the fusion gene contains all the regulatory elements required for vimentin-specific expression. Immunohistochemical staining of fibroblast cultures derived from the transgenic mice with antibodies specific for vimentin and desmin demonstrated that the pVVim2 protein is assembled into filaments that co-localize with the endogenous vimentin filaments. The expression of pVVim2 protein in mesenchymal cells does not interfere with the function of vimentin in these cells.
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- Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
- Bloemendal H., Lenstra J. A., Dodemont H., Ramaekers F. C., Groeneveld A. A., Dunia I., Benedetti E. L. SV40-transformed hamster lens epithelial cells: a novel system for the isolation of cytoskeletal messenger RNAs and their translation products. Exp Eye Res. 1980 Nov;31(5):513–525. doi: 10.1016/s0014-4835(80)80010-8. [DOI] [PubMed] [Google Scholar]
- Broers J. L., Carney D. N., Klein Rot M., Schaart G., Lane E. B., Vooijs G. P., Ramaekers F. C. Intermediate filament proteins in classic and variant types of small cell lung carcinoma cell lines: a biochemical and immunochemical analysis using a panel of monoclonal and polyclonal antibodies. J Cell Sci. 1986 Jul;83:37–60. doi: 10.1242/jcs.83.1.37. [DOI] [PubMed] [Google Scholar]
- Cuypers H. T., Selten G., Quint W., Zijlstra M., Maandag E. R., Boelens W., van Wezenbeek P., Melief C., Berns A. Murine leukemia virus-induced T-cell lymphomagenesis: integration of proviruses in a distinct chromosomal region. Cell. 1984 May;37(1):141–150. doi: 10.1016/0092-8674(84)90309-x. [DOI] [PubMed] [Google Scholar]
- Fisher D. Z., Chaudhary N., Blobel G. cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6450–6454. doi: 10.1073/pnas.83.17.6450. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Schiller D. L., Winter S., Jarasch E. D., Moll R., Denk H., Jackson B. W., Illmensee K. Differentiation-related patterns of expression of proteins of intermediate-size filaments in tissues and cultured cells. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):431–453. doi: 10.1101/sqb.1982.046.01.041. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Schmid E., Winter S., Osborn M., Weber K. Widespread occurrence of intermediate-sized filaments of the vimentin-type in cultured cells from diverse vertebrates. Exp Cell Res. 1979 Oct 1;123(1):25–46. doi: 10.1016/0014-4827(79)90418-x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Lazarides E. Intermediate filaments: a chemically heterogeneous, developmentally regulated class of proteins. Annu Rev Biochem. 1982;51:219–250. doi: 10.1146/annurev.bi.51.070182.001251. [DOI] [PubMed] [Google Scholar]
- McKeon F. D., Kirschner M. W., Caput D. Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins. Nature. 1986 Feb 6;319(6053):463–468. doi: 10.1038/319463a0. [DOI] [PubMed] [Google Scholar]
- Ngai J., Bond V. C., Wold B. J., Lazarides E. Expression of transfected vimentin genes in differentiating murine erythroleukemia cells reveals divergent cis-acting regulation of avian and mammalian vimentin sequences. Mol Cell Biol. 1987 Nov;7(11):3955–3970. doi: 10.1128/mcb.7.11.3955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborn M., Weber K. Intermediate filaments: cell-type-specific markers in differentiation and pathology. Cell. 1982 Dec;31(2 Pt 1):303–306. doi: 10.1016/0092-8674(82)90122-2. [DOI] [PubMed] [Google Scholar]
- Pieper F. R., Slobbe R. L., Ramaekers F. C., Cuypers H. T., Bloemendal H. Upstream regions of the hamster desmin and vimentin genes regulate expression during in vitro myogenesis. EMBO J. 1987 Dec 1;6(12):3611–3618. doi: 10.1002/j.1460-2075.1987.tb02692.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quax-Jeuken Y. E., Quax W. J., Bloemendal H. Primary and secondary structure of hamster vimentin predicted from the nucleotide sequence. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3548–3552. doi: 10.1073/pnas.80.12.3548. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Quax W., van den Heuvel R., Egberts W. V., Quax-Jeuken Y., Bloemendal H. Intermediate filament cDNAs from BHK-21 cells: demonstration of distinct genes for desmin and vimentin in all vertebrate classes. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5970–5974. doi: 10.1073/pnas.81.19.5970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quinlan R. A., Franke W. W. Heteropolymer filaments of vimentin and desmin in vascular smooth muscle tissue and cultured baby hamster kidney cells demonstrated by chemical crosslinking. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3452–3456. doi: 10.1073/pnas.79.11.3452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ramaekers F. C., Osborn M., Schimid E., Weber K., Bloemendal H., Franke W. W. Identification of the cytoskeletal proteins in lens-forming cells, a special epitheloid cell type. Exp Cell Res. 1980 Jun;127(2):309–327. doi: 10.1016/0014-4827(80)90437-1. [DOI] [PubMed] [Google Scholar]
- Ramaekers F. C., Puts J. J., Moesker O., Kant A., Huysmans A., Haag D., Jap P. H., Herman C. J., Vooijs G. P. Antibodies to intermediate filament proteins in the immunohistochemical identification of human tumours: an overview. Histochem J. 1983 Jul;15(7):691–713. doi: 10.1007/BF01002988. [DOI] [PubMed] [Google Scholar]
- Ramaekers F. C., Verheijen R. H., Moesker O., Kant A., Vooijs G. P., Herman C. J. Mesodermal mixed tumor. Diagnosis by analysis of intermediate filament proteins. Am J Surg Pathol. 1983 Jun;7(4):381–385. [PubMed] [Google Scholar]
- Rittling S. R., Baserga R. Functional analysis and growth factor regulation of the human vimentin promoter. Mol Cell Biol. 1987 Nov;7(11):3908–3915. doi: 10.1128/mcb.7.11.3908. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinert P. M., Idler W. W., Cabral F., Gottesman M. M., Goldman R. D. In vitro assembly of homopolymer and copolymer filaments from intermediate filament subunits of muscle and fibroblastic cells. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3692–3696. doi: 10.1073/pnas.78.6.3692. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinert P. M., Steven A. C., Roop D. R. The molecular biology of intermediate filaments. Cell. 1985 Sep;42(2):411–420. doi: 10.1016/0092-8674(85)90098-4. [DOI] [PubMed] [Google Scholar]
- 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]
- Virtanen I., Lehto V. P., Lehtonen E., Vartio T., Stenman S., Kurki P., Wager O., Small J. V., Dahl D., Badley R. A. Expression of intermediate filaments in cultured cells. J Cell Sci. 1981 Aug;50:45–63. doi: 10.1242/jcs.50.1.45. [DOI] [PubMed] [Google Scholar]
- 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]