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A new troponin T and cDNA clones for 13 different muscle proteins, found by shotgun sequencing

Nature volume 302, pages 718–721 (1983)Cite this article

Abstract

Complete amino acid sequences have been established for 19 muscle-related proteins and these proteins are each sufficiently abundant to suggest that their mRNA levels are about 0.4% or higher. Based on these considerations, a simple theoretical analysis shows that clones for most of these proteins can be identified within a complementary DNA library by sequencing cDNA inserts from 150–200 randomly selected clones. This procedure should not only rigorously identify specific clones, but it could also uncover amino acid sequence variants of major muscle proteins such as the troponins1–6. We have determined sequences for about 20,000 nucleotides within 178 randomly selected clones of a rabbit muscle cDNA library, and report here that in addition to finding sequences encoding the two known skeletal muscle isotypes of troponin C7–9, we have discovered sequences encoding two forms of troponin T. Over the region of nucleotide sequence overlap in the troponin T clones, the new isotype diverges significantly from its counterpart10. Altogether, clones for 13 of the 19 known muscle-specific proteins were identified, in addition to the clone for the new troponin T isotype.

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References

  1. Dhoot, G. K. & Perry, S.V. Nature 278, 714 (1979).

    Article  ADS  CAS  Google Scholar 

  2. Hitchcock, S. E. Biochemistry 12, 2509–2515 (1973).

    Article  CAS  Google Scholar 

  3. Perry, S. V. & Cole, H. A. Biochem. J. 141, 733–743 (1974).

    Article  CAS  Google Scholar 

  4. Cole, H. A. & Perry, S. V. Biochem. J. 149, 525–533 (1975).

    Article  CAS  Google Scholar 

  5. Wilkinson, J. M. Biochem. J. 169, 229–238 (1978).

    Article  CAS  Google Scholar 

  6. van Eerd, J. P. & Takahashi, K. Biochem. biophys. Res. Commun. 64, 122–127 (1975).

    Article  CAS  Google Scholar 

  7. Collins, J. H., Potter, J. D., Horn, M. H., Wilshire, G. & Jackman, N. FEBS Lett. 36, 268–272 (1973).

    Article  CAS  Google Scholar 

  8. Collins, J. H. Biochem. biophys. Res. Commun. 58, 301–308 (1974).

    Article  CAS  Google Scholar 

  9. Wilkinson, J. M. Eur. J. Biochem. 103, 179–188 (1980).

    Article  CAS  Google Scholar 

  10. Pearlstone, J. R., Carpenter, M. R., Johnson, P. & Smillie, L. B. Proc. natn. Acad. Sci. U.S.A. 73, 1902–1906 (1976).

    Article  ADS  CAS  Google Scholar 

  11. Messing, J. in 3rd Cleveland Symp. Macromolecules: Recombinant DNA (ed. Walton, A.) 143–153 (Elsevier, Amsterdam, 1981).

    Google Scholar 

  12. Romero-Herrera, A. E., Lehemann, H. & Castillo, O. Biochim. biophys. Acta 439, 51–54 (1976).

    Article  CAS  Google Scholar 

  13. Frank, G. & Weeds, A. G. Eur. J. Biochem. 44, 317–334 (1974).

    Article  CAS  Google Scholar 

  14. Weeds, A. G. FEBS Lett. 59, 203–208 (1975).

    Article  CAS  Google Scholar 

  15. Enfield, D. L., Ericsson, L. H., Blu, H. E., Fischer, E. H. & Neurath, M. Proc. natn. Acad. Sci. U.S.A. 72, 1309–1313 (1975).

    Article  ADS  CAS  Google Scholar 

  16. Capony, J.-P., Pina, C. & Pechere, J.-F. Eur. J. Biochem. 70, 123–135 (1976).

    Article  CAS  Google Scholar 

  17. Kiltz, H.-H., Keil, W., Griesbach, M., Petry, K. & Meyer, H. Hoppe-Seyler's Z. physiol. Chem. 358, 123–127 (1977).

    Article  CAS  Google Scholar 

  18. AlLen, G. Biochem. J. 187, 545–563, 565–575 (1980).

    Article  CAS  Google Scholar 

  19. AlLen, G., Bottomley, R. C. & Trinnaman, B. J. Biochem. J. 187, 577–589 (1980).

    Article  CAS  Google Scholar 

  20. AlLen, G., Trinnaman, B. J. & Green, N. M. Biochem. J. 187, 591–616 (1980).

    Article  CAS  Google Scholar 

  21. Schwartz, R. J., & Rothblum, K. Biochemistry 19, 2506–2514 (1980).

    Article  CAS  Google Scholar 

  22. Arnold, H.-H. & Siddiqui, M. A. Q. Biochemistry 18, 647–654 (1979).

    Article  CAS  Google Scholar 

  23. Benoff, S. & Nadal-Ginard, B. Proc. natn. Acad. Sci. U.S.A. 76, 1853–1857 (1979).

    Article  ADS  CAS  Google Scholar 

  24. Ebashi, E. & Nonomura, Y. in The Structure and Function of Muscle (ed. Bovone, G. J.) 285–362 (Academic, New York, 1973).

    Google Scholar 

  25. Hastings, K. E. M. & Emerson, C. P. Proc. natn. Acad. Sci. U.S.A. 79, 1553–1557 (1982).

    Article  ADS  CAS  Google Scholar 

  26. Ordahl, C. P., Kioussis, D., Tilghman, S. M., Ovitt, C. E. & Fornwald, J. Proc. natn. Acad. Sci. U.S.A. 77, 4519–4523 (1980).

    Article  ADS  CAS  Google Scholar 

  27. Robbins, J., Freyer, G. A., Chisholm, D. & Gilliam, T. C. J. biol. Chem. 257, 549–556 (1982).

    CAS  PubMed  Google Scholar 

  28. Katcoff, D. et al. Proc. natn. Acad. Sci. U.S.A. 77, 960–964 (1980).

    Article  ADS  CAS  Google Scholar 

  29. Sinha, A. M. et al. Proc. natn. Acad. Sci. U.S.A. 79, 5847–5851 (1982).

    Article  ADS  CAS  Google Scholar 

  30. Lemischka, I. R., Farmer, S., Racaniello, V. R. & Sharp, P. A. J. molec. Biol. 151, 101 (1981).

    Article  CAS  Google Scholar 

  31. Aviv, H. & Leder, P. Proc. natn. Acad. Sci. U.S.A. 69, 1408–1412 (1972).

    Article  ADS  CAS  Google Scholar 

  32. Crabtree, G. R. & Kant, J. A. J. biol. Chem. 256, 9718–9723 (1981).

    CAS  PubMed  Google Scholar 

  33. Roberts, T. M. & Lauer, G. D. Meth. Enzym. 68, 473–482 (1979).

    Article  CAS  Google Scholar 

  34. Maniatis, T., Jeffrey, A. & van deSande, H. Biochemistry 14, 3787–3794 (1975).

    Article  CAS  Google Scholar 

  35. Maxam, A. & Gilbert, W. Meth. Enzym. 65, 499–560 (1980).

    Article  CAS  Google Scholar 

  36. Messing, J., Crea, R. & Seeburg, P. H. Nucleic Acids Res. 9, 309–321 (1981).

    Article  CAS  Google Scholar 

  37. Sanger, F., Nicklen, S. & Coulson, A. R. Proc. natn. Acad. Sci. U.S.A. 74, 5463–5467 (1977).

    Article  ADS  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139, USA

    Scott D. Putney & Paul Schimmel

  2. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts, 02139, USA

    Walter C. Herlihy

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  1. Scott D. Putney
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  2. Walter C. Herlihy
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  3. Paul Schimmel
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Putney, S., Herlihy, W. & Schimmel, P. A new troponin T and cDNA clones for 13 different muscle proteins, found by shotgun sequencing. Nature 302, 718–721 (1983). https://doi.org/10.1038/302718a0

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