A role for the proteasome in the light response of the timeless clock protein

Science

Volumn 285, Issue 5434, 1999, Pages 1737-1741

  Naidoo, Nirinjini ^a   Song, Wei ^a   Hunter Ensor, Melissa ^a,b   Sehgal, Amita ^a  

^a UNIVERSITY OF PENNSYLVANIA   (United States)

^b MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PROTEASOME; REGULATOR PROTEIN;

ARTICLE; BIOLOGICAL RHYTHM; CIRCADIAN RHYTHM; DROSOPHILA; ENZYME ACTIVITY; NONHUMAN; PRIORITY JOURNAL; PROTEIN DEGRADATION; PROTEIN PHOSPHORYLATION;

ACETYLCYSTEINE; ANIMALS; BIOLOGICAL CLOCKS; CELLS, CULTURED; CIRCADIAN RHYTHM; CYSTEINE ENDOPEPTIDASES; CYSTEINE PROTEINASE INHIBITORS; DARKNESS; DROSOPHILA; DROSOPHILA PROTEINS; FEEDBACK; INSECT PROTEINS; LEUCINE; LEUPEPTINS; LIGHT; MULTIENZYME COMPLEXES; NEURONS; PHOSPHORYLATION; PHOSPHOTYROSINE; PROTEASE INHIBITORS; PROTEASOME ENDOPEPTIDASE COMPLEX; UBIQUITINS;

EID: 0033543596     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5434.1737     Document Type: Article

References (41)

1. P. E. Hardin and A. Sehgal, in Handbook of Behavioral State Control: Cellular and Molecular Mechanisms (CRC Press, Boca Raton, FL, 1999), pp. 61-74.
2. J. C. Dunlap, Cell 96, 271 (1999).
3. S. B. Marrus, H. Zeng, M. Rosbash, EMBO J. 15, 6877 (1996).
4. M. E. Dembinska, R. Stanewsky, J. C. Hall, M. Rosbash, J. Biol. Rhythms 12, 157 (1997).
5. Y. Chen, M. Hunter-Ensor, P. Schotland, A. Sehgal, ibid. 13, 364 (1998).
6. J. L Price et al., Cell 94, 83 (1998).
7. M. Hunter-Ensor, A. Ousley, A. Sehgal, ibid. 84, 677 (1996).
8. C. Lee, V. Parikh, T. Itsukaichi, K. Bae, I. Edery, Science 271, 1740 (1996).
9. M. P. Myers, K. Wager-Smith, A. Rothenflugh, M. W. Young, ibid., p. 1736.
10. H. Zeng, Z. Qian, M. P. Myers, M. Rosbash, Nature 380, 129 (1996).
11. Z. Yang, M. Emerson, H. S. Su, A. Sehgal, Neuron 21, 215 (1998).
12. V. Suri, Q. Zuwei, J. C. Hall, M. Rosbash, ibid., p. 225.
13. N. Naidoo, W. Song, M. Hunter-Ensor, A. Sehgal, data not shown.
14. K. L. Rock et al., Cell 78, 761 (1994).
15. E. Carafoli and M. Molinari, Biochem. Biophys. Res. Commun. 247, 193 (1998).
16. In about 35% of the experiments, the pulsed extract did not display proteolytic activity or resulted in incomplete proteotysis, yielding a fragment with a molecular mass (MW) between 66 and 97 kD. We regarded complete or partial disappearance of the lower of two bands (which are recognized by the antibody to TIM) as positive. We usually also observed reduction in the upper band, which frequently shifted down in molecular mass.
17. G. Fenteany et al., Science 268, 726 (1995).
18. A. Craiu et al., J. Biol. Chem. 272, 13437 (1997).
19. M. Kaneko, C. Helfrich-Forster, J. C. Hall, J. Neurosci. 17, 6745 (1997); I. F. Emery, J. M. Noveral, C. F. Jamison, K. K. Siwicki, Proc. Natl. Acad. Sci. U.S.A. 94, 4092 (1997).
20. M. Kaneko, C. Helfrich-Forster, J. C. Hall, J. Neurosci. 17, 6745 (1997); I. F. Emery, J. M. Noveral, C. F. Jamison, K. K. Siwicki, Proc. Natl. Acad. Sci. U.S.A. 94, 4092 (1997).
21. W. Baumeister, J. Walz, F. Zuhl, E. Seemuller, Cell 92, 367 (1998).
22. J. A. Gastel, P. H. Roseboom, P. A. Rinaldi, J. L. Weller, D. C. Klein, Science 279, 1358 (1998).
23. E. Geier et al., ibid. 283, 978 (1999).
24. M. Rechsteiner and S. W. Rogers, Trends Biochem. Sci. 21, 267 (1996).
25. C. Pickart, FASEB J. 11, 1055 (1997).
26. M. Treier, L. M. Staszewski, D. Bohmann, Cell 78, 787 (1994).
27. I. V. Davydov, D. Patra, A. Varshavsky, Arch. Biochem. Biophys. 357, 317 (1998).
28. M. Vecchi and G. Carpenter, J. Cell Biol. 139, 995 (1997).
29. R. J. Deshaies, V. Chau, M. W. Kirschner, EMBO J. 14, 303 (1995).
30. J. Yaglom et al., Mol. Cell. Biol. 15, 731 (1995).
31. S. Lahav-Baratz, V. Sudakin, J. V. Ruderman, A. Hershko, Proc. Natl. Acad. Sci. U.S.A. 92, 9303 (1995).
32. J. F. Geissler et al., J. Biol. Chem. 265, 22255 (1990).
33. T. Tamaoki et al., Biochem. Biophys. Res. Commun. 135, 397 (1986).
34. E. Kobayashi et al., J. Antibiotics 42, 1470 (1989).
35. D. Dudley, L. Pang, S. Decker, A. Bridges, A. Saltiel, Proc. Natl. Acad. Sci. U.S.A. 92, 7686 (1995).
36. D. Alessi, A. Cuenda, P. Cohen, D. Dudley, A. Saltiel, J. Biol. Chem. 270, 27489 (1995).
37. Heads from yellow-white flies collected at ZT19 or ZT20 were homogenized in IP buffer [10 mM Hepes, 150 mM NaCl, and 1 mM EDTA (pH 7.5)]. Triton X-100 was added to the lysate at a final concentration of 1% before incubation at 4°C for 30 min. The lysate was centrifuged at 5000 rpm. The protein concentration of the supernatant was determined before immunoprecipitation by means of the Bio-Rad assay.
38. The head extract was made by homogenizing fly heads in IP buffer, adding Triton X-100 to a final concentration of 1% and incubating the lysate at 4°C for 30 min with rotation. The lysate was centrifuged at 500 to 700 rpm at 4°C before use in the assay.
39. After light treatment, the tissue was fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 30 min, rinsed in PBS containing 1% Triton for 2 periods of 20 min, and then blocked in PBS containing 0.5% Triton with 10% normal goat serum for 1 hour. Primary antibodies (1:1500 dilution of anti-TIM and a 1:10 000 dilution of rabbit anti-PDH) were incubated overnight at 4°C in blocking solution. The following day, the tissue was washed three times, for 1 hour each time, in rinse solution and incubated for 1 hour with a 1:500 dilution of fluorescein isothiocyanate-conjugated goat antibody to rabbit (Jackson Immunoresearch) and a 1:1000 dilution of Cy3-conjugated donkey antibody to rat (Jackson Immunoresearch). The tissue was again washed with rinse solution, then mounted with Vectastain H 1000 medium with 4′,6′-diamidino-2-phenylindole. Slides were analyzed with a Leica DMRE microscope.
40. L. Saez and M. W. Young, Neuron 17, 911 (1996).
41. We thank C. Pickart and J. Field for critical reading of the manuscript and for many useful suggestions, L. Matthews and K. Siwicki for demonstrating the larval dissections, K. Hu for sharing results of her experiments, and other members of the laboratory for general advice. Supported by grants from NIH and NSF to A.S. M.H.E. was supported by an NIH training grant to the Behavioral Neuroscience program. A.S. is an assistant investigator in the Howard Hughes Medical Institute.