Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1988 Dec;8(12):5561–5565. doi: 10.1128/mcb.8.12.5561

Isolation and characterization of a cDNA that encodes mouse fibroblast tropomyosin isoform 2.

K Takenaga 1, Y Nakamura 1, K Tokunaga 1, H Kageyama 1, S Sakiyama 1
PMCID: PMC365662  PMID: 3244365

Abstract

We isolated and characterized a cDNA clone encoding tropomyosin isoform 2 (TM2) from a mouse fibroblast cDNA library. TM2 was found to contain 284 amino acids and was closely related to the rat smooth and skeletal muscle alpha-TMs and the human fibroblast TM3. The amino acid sequence of TM2 showed a nearly complete match with that of human fibroblast TM3 except for the region from amino acids 189 to 213, the sequence of which was identical to those of rat smooth and skeletal muscle alpha-TMs. These results suggest that TM2 is expressed from the same gene that encodes the smooth muscle alpha-TM, the skeletal muscle alpha-TM, and TM3 via an alternative RNA-splicing mechanism. Comparison of the expression of TM2 mRNA in low-metastatic Lewis lung carcinoma P29 cells and high-metastatic D6 cells revealed that it was significantly less in D6 cells than in P29 cells, supporting our previous observations (K. Takenaga, Y. Nakamura, and S. Sakiyama, Mol. Cell. Biol. 8:3934-3937, 1988) at the protein level.

Full text

PDF
5561

Images in this article

5562Image
on p.5562
5564Image
on p.5564
5564Image
on p.5564

Selected References

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

  1. Bernstein B. W., Bamburg J. R. Tropomyosin binding to F-actin protects the F-actin from disassembly by brain actin-depolymerizing factor (ADF). Cell Motil. 1982;2(1):1–8. doi: 10.1002/cm.970020102. [DOI] [PubMed] [Google Scholar]
  2. Cooper H. L., Feuerstein N., Noda M., Bassin R. H. Suppression of tropomyosin synthesis, a common biochemical feature of oncogenesis by structurally diverse retroviral oncogenes. Mol Cell Biol. 1985 May;5(5):972–983. doi: 10.1128/mcb.5.5.972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fattoum A., Hartwig J. H., Stossel T. P. Isolation and some structural and functional properties of macrophage tropomyosin. Biochemistry. 1983 Mar 1;22(5):1187–1193. doi: 10.1021/bi00274a031. [DOI] [PubMed] [Google Scholar]
  4. Hendricks M., Weintraub H. Multiple tropomyosin polypeptides in chicken embryo fibroblasts: differential repression of transcription by Rous sarcoma virus transformation. Mol Cell Biol. 1984 Sep;4(9):1823–1833. doi: 10.1128/mcb.4.9.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hendricks M., Weintraub H. Tropomyosin is decreased in transformed cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5633–5637. doi: 10.1073/pnas.78.9.5633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hitchcock S. E., Huxley H. E., Szent-Györgyi A. G. Calcium sensitive binding of troponin to actin-tropomyosin: a two-site model for troponin action. J Mol Biol. 1973 Nov 15;80(4):825–836. doi: 10.1016/0022-2836(73)90212-x. [DOI] [PubMed] [Google Scholar]
  7. Lazarides E. Tropomyosin antibody: the specific localization of tropomyosin in nonmuscle cells. J Cell Biol. 1975 Jun;65(3):549–561. doi: 10.1083/jcb.65.3.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Leavitt J., Latter G., Lutomski L., Goldstein D., Burbeck S. Tropomyosin isoform switching in tumorigenic human fibroblasts. Mol Cell Biol. 1986 Jul;6(7):2721–2726. doi: 10.1128/mcb.6.7.2721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lin C. S., Leavitt J. Cloning and characterization of a cDNA encoding transformation-sensitive tropomyosin isoform 3 from tumorigenic human fibroblasts. Mol Cell Biol. 1988 Jan;8(1):160–168. doi: 10.1128/mcb.8.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MacLeod A. R., Gooding C. Human hTM alpha gene: expression in muscle and nonmuscle tissue. Mol Cell Biol. 1988 Jan;8(1):433–440. doi: 10.1128/mcb.8.1.433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MacLeod A. R., Houlker C., Reinach F. C., Smillie L. B., Talbot K., Modi G., Walsh F. S. A muscle-type tropomyosin in human fibroblasts: evidence for expression by an alternative RNA splicing mechanism. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7835–7839. doi: 10.1073/pnas.82.23.7835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Matsumura F., Lin J. J., Yamashiro-Matsumura S., Thomas G. P., Topp W. C. Differential expression of tropomyosin forms in the microfilaments isolated from normal and transformed rat cultured cells. J Biol Chem. 1983 Nov 25;258(22):13954–13964. [PubMed] [Google Scholar]
  13. Matsumura F., Yamashiro-Matsumura S., Lin J. J. Isolation and characterization of tropomyosin-containing microfilaments from cultured cells. J Biol Chem. 1983 May 25;258(10):6636–6644. [PubMed] [Google Scholar]
  14. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  16. Ruiz-Opazo N., Nadal-Ginard B. Alpha-tropomyosin gene organization. Alternative splicing of duplicated isotype-specific exons accounts for the production of smooth and striated muscle isoforms. J Biol Chem. 1987 Apr 5;262(10):4755–4765. [PubMed] [Google Scholar]
  17. Ruiz-Opazo N., Weinberger J., Nadal-Ginard B. Comparison of alpha-tropomyosin sequences from smooth and striated muscle. Nature. 1985 May 2;315(6014):67–70. doi: 10.1038/315067a0. [DOI] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Takenaga K. Modification of the metastatic potential of tumor cells by drugs. Cancer Metastasis Rev. 1986;5(2):67–75. doi: 10.1007/BF00046423. [DOI] [PubMed] [Google Scholar]
  20. Takenaga K., Nakamura Y., Sakiyama S. Differential expression of a tropomyosin isoform in low- and high-metastatic Lewis lung carcinoma cells. Mol Cell Biol. 1988 Sep;8(9):3934–3937. doi: 10.1128/mcb.8.9.3934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Talbot K., MacLeod A. R. Novel form of non-muscle tropomyosin in human fibroblasts. J Mol Biol. 1983 Feb 15;164(1):159–174. doi: 10.1016/0022-2836(83)90091-8. [DOI] [PubMed] [Google Scholar]
  22. Tokunaga K., Takeda K., Kamiyama K., Kageyama H., Takenaga K., Sakiyama S. Isolation of cDNA clones for mouse cytoskeletal gamma-actin and differential expression of cytoskeletal actin mRNAs in mouse cells. Mol Cell Biol. 1988 Sep;8(9):3929–3933. doi: 10.1128/mcb.8.9.3929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tokunaga K., Taniguchi H., Yoda K., Shimizu M., Sakiyama S. Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA. Nucleic Acids Res. 1986 Mar 25;14(6):2829–2829. doi: 10.1093/nar/14.6.2829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wieczorek D. F., Smith C. W., Nadal-Ginard B. The rat alpha-tropomyosin gene generates a minimum of six different mRNAs coding for striated, smooth, and nonmuscle isoforms by alternative splicing. Mol Cell Biol. 1988 Feb;8(2):679–694. doi: 10.1128/mcb.8.2.679. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES