Force generation of organelle transport measured in vivo by an infrared laser trap

Abstract

ORGANELLE transport along microtubules is believed to be mediated by organelle-associated force-generating molecules1. Two classes of microtubule-based organelle motors have been identified: kinesin2–7 and cytoplasmic dynein8–12. To correlate the mechanochemical basis of force generation with the in vivo behaviour of organelles, it is important to quantify the force needed to propel an organelle along microtubules and to determine the force generated by a single motor molecule. Measurements of force generation are possible under selected conditions in vitro (for example, see refs 13 and 14), but are much more difficult using intact or reactivated cells. Here we combine a useful model system for the study of organelle transport, the giant amoeba Reticulomyxa15, with a novel technique for the non-invasive manipulation of and force application to subcellular components, which is based on a gradient-force optical trap, also referred to as 'optical tweezers'16–19. We demonstrate the feasibility of using controlled manipulation of actively translocating organelles to measure direct force. We have determined the force driving a single organelle along microtubules, allowing us to estimate the force generated by a single motor to be 2.6 × 10-7 dynes.

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References

  1. 1

    Warner, F. D. & McIntosh, J. R. (eds) in Cell Movement Vol. 2 (Liss, New York, 1989).

  2. 2

    Brady, S. T. Nature 317, 73–75 (1985).

    ADS  CAS  Article  Google Scholar 

  3. 3

    Vale, R. D., Reese, T. S. & Sheetz, M. P. Cell 42, 39–50 (1985).

    CAS  Article  Google Scholar 

  4. 4

    Scholey, J. M., Porter, M. E., Grissom, P. M. & McIntosh, J. R. Nature 318, 483–486 (1985).

    ADS  CAS  Article  Google Scholar 

  5. 5

    Schroer, T. A., Schnapp, B. J., Reese, T. S. & Scheetz, M. P. J. Cell Biol. 107, 1785–1792 (1988).

    CAS  Article  Google Scholar 

  6. 6

    Pfister, K. K., Wagner, M. C., Stenoien, D. L., Brady, S. T. & Bloom, G. S. J. Cell Biol. 108, 1453–1463 (1988).

    Article  Google Scholar 

  7. 7

    Brady, S. T., Pfister, K. K. & Bloom, G. S. Proc. natn Acad. Sci. U.S.A. 87, 1061–1065 (1990).

    ADS  CAS  Article  Google Scholar 

  8. 8

    Paschal, B. M., Shpetner, H. S. & Vallee, R. B. J. Cell Biol. 105, 1273–1282 (1987).

    CAS  Article  Google Scholar 

  9. 9

    Paschal, B. M. & Vallee, R. B. Nature 330, 181–183 (1988).

    ADS  Article  Google Scholar 

  10. 10

    Lye, R. J., Porter, M. E., Scholey, J. M. & McIntosh, J. R. Cell 51, 309–318 (1988).

    Article  Google Scholar 

  11. 11

    Euteneuer, U., Koonce, M. P., Pfister, K. K. & Schliwa, M. Nature 332, 176–178 (1988).

    ADS  CAS  Article  Google Scholar 

  12. 12

    Shroer, T. A., Steuer, E. & Sheetz, M. P. Cell 56, 937–946 (1989).

    Article  Google Scholar 

  13. 13

    Kishino, A. & Yanagida, T. Nature 334, 74–76 (1988).

    ADS  CAS  Article  Google Scholar 

  14. 14

    Howard, J., Hudspeth, A. J. & Vale, R. D. Nature 342, 154–158 (1989).

    ADS  CAS  Article  Google Scholar 

  15. 15

    Euteneuer, U. et al. in Cell Movement Vol. 2 (eds Warner, F. D. & McIntosh, J. R.) 155–167 (Liss, New York, 1989).

    Google Scholar 

  16. 16

    Ashkin, A. & Dziedzic, J. M. Science 235, 1517–1520 (1987).

    ADS  CAS  Article  Google Scholar 

  17. 17

    Ashkin, A. & Dziedzic, J. M. & Yamane, T. Nature 330, 769–771 (1987).

    ADS  CAS  Article  Google Scholar 

  18. 18

    Ashkin, A. & Dziedzic, J. M. Proc. natn. Acad. Sci. U.S.A. 86, 7914–7918 (1989).

    ADS  CAS  Article  Google Scholar 

  19. 19

    Block, S. M., Blair, D. F. & Berg, H. C. Nature 338, 514–517 (1989).

    ADS  CAS  Article  Google Scholar 

  20. 20

    Koonce, M. P. & Schliwa, M. J. Cell Biol. 103, 605–612 (1986).

    CAS  Article  Google Scholar 

  21. 21

    Euteneuer, U., Johnson, K. B. & Schliwa, M. Eur. J. Cell Biol. 50, 34–40 (1989).

    Google Scholar 

  22. 22

    Schliwa, M., Shimizu, T., Vale, R. D. & Euteneuer, U. J. Cell Biol. (in the press).

  23. 23

    Ashkin, A., Dziedzic, J. M., Bjorkholm, J. E. & Chu, S. Optics Lett. 11, 288–290 (1986).

    ADS  CAS  Article  Google Scholar 

  24. 24

    Koonce, M. P. & Schliwa, M. J. Cell Biol. 100, 322–326 (1985).

    CAS  Article  Google Scholar 

  25. 25

    Kamimura, S. & Takahashi, K. Nature 293, 566–568 (1981).

    ADS  CAS  Article  Google Scholar 

  26. 26

    Oiwa, K. & Takahashi, K. Cell Struct. Funct. 13, 193–205 (1988).

    CAS  Article  Google Scholar 

  27. 27

    Hiramoto, Y. in Cilia and Flagella (ed. Sleigh, M. A.) 177–196 (Academic, London, 1974).

    Google Scholar 

  28. 28

    Brokaw, C. J. in Molecules and Cell Movement (eds Inoue, S. & Stephens, R. E.) 165–179 (Raven, New York, 1975).

    Google Scholar 

  29. 29

    Block, S., Goldstein, L. S. B. & Schnapp, B. J. J. Cell Biol. 109, 81a (1989).

    Google Scholar 

  30. 30

    Kishino, A. & Yanagida, T. Nature 334, 74–76 (1988).

    ADS  CAS  Article  Google Scholar 

  31. 31

    Oosawa, F. Biorheology 14, 11–19 (1977).

    CAS  Article  Google Scholar 

  32. 32

    Schliwa, M. & van Blerkom, J. J. Cell Biol. 90, 222–235 (1981).

    CAS  Article  Google Scholar 

  33. 33

    Euteneur, U., Haimo, L. T. & Schliwa, M. Eur. J. Cell Biol. 49, 373–376 (1989).

    Google Scholar 

  34. 34

    Koonce, M. P., Euteneuer, V., McDonald, K. L., Menzel, D. & Schiwa, M. Cell Motil. Cytoskel. 6, 521–533 (1986).

    CAS  Article  Google Scholar 

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Ashkin, A., Schütze, K., Dziedzic, J. et al. Force generation of organelle transport measured in vivo by an infrared laser trap. Nature 348, 346–348 (1990). https://doi.org/10.1038/348346a0

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