Drosophila S6 kinase: A regulator of cell size

Science

Volumn 285, Issue 5436, 1999, Pages 2126-2129

  Montagne, Jacques ^a   Stewart, Mary J ^a,c   Stocker, Hugo ^b   Hafen, Ernst ^b   Kozma, Sara C ^a   Thomas, George ^a  

^a FRIEDRICH MIESCHER INSTITUTE FOR BIOMEDICAL RESEARCH   (Switzerland)

^b UNIVERSITY OF ZURICH   (Switzerland)

^c NORTH DAKOTA STATE UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

GENE PRODUCT; MUTANT PROTEIN; PHOSPHOTRANSFERASE; RIBOSOME PROTEIN;

ARTICLE; BODY SIZE; CELL COUNT; CELL GROWTH; CELL PROLIFERATION; CELL SIZE; DNA SEQUENCE; DROSOPHILA; GENE MUTATION; GROWTH REGULATION; NONHUMAN; NORTHERN BLOTTING; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION;

ANIMALS; BASE SEQUENCE; BODY CONSTITUTION; CELL COUNT; CELL DIVISION; CELL SIZE; DROSOPHILA MELANOGASTER; EPITHELIAL CELLS; FEMALE; GENES, INSECT; LARVA; MALE; METAMORPHOSIS, BIOLOGICAL; MOLECULAR SEQUENCE DATA; MUTATION; RIBOSOMAL PROTEIN S6 KINASES; WING;

EID: 0032843917     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5436.2126     Document Type: Article

References (46)

1. J. M. Mitchison, The Biology of the Cell Cycle (Cambridge Univ. Press, Cambridge, 1971); T. T. Su and P. H. O'Farrell, Curr. Biol. 8, R687 (1998); I. Conlon and M. Raff, Cell 96, 235 (1999).
2. J. M. Mitchison, The Biology of the Cell Cycle (Cambridge Univ. Press, Cambridge, 1971); T. T. Su and P. H. O'Farrell, Curr. Biol. 8, R687 (1998); I. Conlon and M. Raff, Cell 96, 235 (1999).
3. J. M. Mitchison, The Biology of the Cell Cycle (Cambridge Univ. Press, Cambridge, 1971); T. T. Su and P. H. O'Farrell, Curr. Biol. 8, R687 (1998); I. Conlon and M. Raff, Cell 96, 235 (1999).
4. G. C. Johnston, J. R. Pringle, L. H. Hartwell, Exp. Cell Res. 105, 79 (1977); P. Nurse, in The Evolution of Genome Size, T. Cavalier-Smith, Ed. (Wiley, New York, 1985), pp. 185-196; K. Nasmyth, Curr. Opin. Cell Biol. 5, 166 (1993).
5. G. C. Johnston, J. R. Pringle, L. H. Hartwell, Exp. Cell Res. 105, 79 (1977); P. Nurse, in The Evolution of Genome Size, T. Cavalier-Smith, Ed. (Wiley, New York, 1985), pp. 185-196; K. Nasmyth, Curr. Opin. Cell Biol. 5, 166 (1993).
6. G. C. Johnston, J. R. Pringle, L. H. Hartwell, Exp. Cell Res. 105, 79 (1977); P. Nurse, in The Evolution of Genome Size, T. Cavalier-Smith, Ed. (Wiley, New York, 1985), pp. 185-196; K. Nasmyth, Curr. Opin. Cell Biol. 5, 166 (1993).
7. T. P. Neufeld, A. F. de la Cruz, L. A. Johnston, B. A. Edgar, Cell 93, 1183 (1998).
8. K. Nasmyth, Nature 382, 28 (1996); G. Thomas and M. N. Hall, Curr. Opin. Cell. Biol. 9, 782 (1997).
9. K. Nasmyth, Nature 382, 28 (1996); G. Thomas and M. N. Hall, Curr. Opin. Cell. Biol. 9, 782 (1997).
10. H. Shima et al., EMBO J. 17, 6649 (1998).
11. H. B. J. Jefferies et al., ibid. 12, 3693 (1997).
12. A. Lambertsson, Adv. Genet. 38, 69 (1998).
13. G. H. Karpen and A. C. Spradling, Genetics 132, 737 (1992); T. F. Consortium, Nucleic Acids Res. 22, 3456 (1994).
14. G. H. Karpen and A. C. Spradling, Genetics 132, 737 (1992); T. F. Consortium, Nucleic Acids Res. 22, 3456 (1994).
15. The gene was mapped to cytological position 64F1-3 as described (26). A collection of P elements in this region were analyzed for restriction fragment length polymorphisms, with one strain, fs(3)07084, showing an alteration when probed on Southern (DNA) blots with the dS6K cDNA.
16. Fly strains were as described [D. L. Lindsley and G. G. Zimm, The Genome of Drosophila melanogaster (Academic Press, San Diego, CA, 1992)] and were maintained under standard conditions. P-element revertants were generated by standard genetic techniques. dS6K and S6K cDNAs were from (27) and (5), respectively. To generate transgenic flies, we transformed a y, w, ac stock as described [G. M. Rubin and A. C. Spradling, Science 218, 348 (1982)] with pBD1119 derivatives that contained either dS6K, S6K1, or S6K2 cDNAs under the control of the α-tubulin promoter [W. E. Theurkauf, H. Baum, P. C. Wensik, Proc. Natl. Acad. Sci. U.S.A. 83, 8477 (1986)]. One copy of the dS6K or S6K transgene rescued the P-insertion or P-excision strains. The two additional Minutes analyzed were M(3)66D and M(2)32A, encoding ribosomal proteins L14 and S13, respectively (7).
17. Fly strains were as described [D. L. Lindsley and G. G. Zimm, The Genome of Drosophila melanogaster (Academic Press, San Diego, CA, 1992)] and were maintained under standard conditions. P-element revertants were generated by standard genetic techniques. dS6K and S6K cDNAs were from (27) and (5), respectively. To generate transgenic flies, we transformed a y, w, ac stock as described [G. M. Rubin and A. C. Spradling, Science 218, 348 (1982)] with pBD1119 derivatives that contained either dS6K, S6K1, or S6K2 cDNAs under the control of the α-tubulin promoter [W. E. Theurkauf, H. Baum, P. C. Wensik, Proc. Natl. Acad. Sci. U.S.A. 83, 8477 (1986)]. One copy of the dS6K or S6K transgene rescued the P-insertion or P-excision strains. The two additional Minutes analyzed were M(3)66D and M(2)32A, encoding ribosomal proteins L14 and S13, respectively (7).
18. Fly strains were as described [D. L. Lindsley and G. G. Zimm, The Genome of Drosophila melanogaster (Academic Press, San Diego, CA, 1992)] and were maintained under standard conditions. P-element revertants were generated by standard genetic techniques. dS6K and S6K cDNAs were from (27) and (5), respectively. To generate transgenic flies, we transformed a y, w, ac stock as described [G. M. Rubin and A. C. Spradling, Science 218, 348 (1982)] with pBD1119 derivatives that contained either dS6K, S6K1, or S6K2 cDNAs under the control of the α-tubulin promoter [W. E. Theurkauf, H. Baum, P. C. Wensik, Proc. Natl. Acad. Sci. U.S.A. 83, 8477 (1986)]. One copy of the dS6K or S6K transgene rescued the P-insertion or P-excision strains. The two additional Minutes analyzed were M(3)66D and M(2)32A, encoding ribosomal proteins L14 and S13, respectively (7).
19. (RNA) blots as described [M. J. Stewart and R. Denell, Mol. Cell. Biol. 13, 2524 (1993)]. Plasmid rescue was as described [V. Pirrotta, in Drosophila, a Practical Approach, D. B. Roberts, Ed. (IRL Press, Oxford, UK, 1986), pp. 82-109]. Chimeric transcripts were reversed-transcribed with avian myeloblastosis virus (Promega) with an oligonucleotide priming from the exon 2 of the dS6K gene. PCR reaction products obtained revealed the presence of a chimeric mRNA containing the P-element sequence and the fusion of exon 1 to exon 2.
20. (RNA) blots as described [M. J. Stewart and R. Denell, Mol. Cell. Biol. 13, 2524 (1993)]. Plasmid rescue was as described [V. Pirrotta, in Drosophila, a Practical Approach, D. B. Roberts, Ed. (IRL Press, Oxford, UK, 1986), pp. 82-109]. Chimeric transcripts were reversed-transcribed with avian myeloblastosis virus (Promega) with an oligonucleotide priming from the exon 2 of the dS6K gene. PCR reaction products obtained revealed the presence of a chimeric mRNA containing the P-element sequence and the fusion of exon 1 to exon 2.
21. 07084 flies. To identify break points, we did PCR amplification from single flies [G. B. Gloor et al., Genetics 135, 81 (1993)], with oligonucleotides priming in the genomic sequence upstream of the P insertion point and in exon 8 of the dS6K gene.
22. 07084 flies. To identify break points, we did PCR amplification from single flies [G. B. Gloor et al., Genetics 135, 81 (1993)], with oligonucleotides priming in the genomic sequence upstream of the P insertion point and in exon 8 of the dS6K gene.
23. 2, located between veins 3 and 4 of the dorsal wing blade, up to the posterior cross vein. The total number of dorsal wing blade intervein cells was calculated by multiplying the number above by the area of the wing blade. For scanning electron microscopy, 3-to 4-day-old flies were anesthetized and immersed in 70% acetone. After critical point drying, they were mounted and coated with gold. The specimens were observed with a Hitachi S-800 field emission electron microscope at 6 kV.
24. M. Schubiger and J. Palka, Dev. Biol. 123, 145 (1987) and (26), pp. 171-179. Female third instar wandering larvae were collected for disc dissection upon immobilization on the wall of the tube. Early prepupae were identified by spiracle eversion and before epidermal hardening. Wing disc cells were analyzed with a flow analysis cytometry sorter as described (3). The proliferation rate was estimated to be reduced by twofold by analyzing developing larvae. Four additional days were required to complete larval development and one additional day for pupation.
25. M. Schubiger and J. Palka, Dev. Biol. 123, 145 (1987) and (26), pp. 171-179. Female third instar wandering larvae were collected for disc dissection upon immobilization on the wall of the tube. Early prepupae were identified by spiracle eversion and before epidermal hardening. Wing disc cells were analyzed with a flow analysis cytometry sorter as described (3). The proliferation rate was estimated to be reduced by twofold by analyzing developing larvae. Four additional days were required to complete larval development and one additional day for pupation.
26. M. Madhavan and H. A. Schneiderman, Wilhelm Roux's Arch. 183, 269 (1977).
27. A. Garcia-Bellido and J. R. Merriam, Dev. Biol. 24, 61 (1971); M. J. Fain and B. Stevens, ibid. 92, 247 (1982); B. J. Graves and G. Schubiger, ibid. 93, 104 (1982).
28. A. Garcia-Bellido and J. R. Merriam, Dev. Biol. 24, 61 (1971); M. J. Fain and B. Stevens, ibid. 92, 247 (1982); B. J. Graves and G. Schubiger, ibid. 93, 104 (1982).
29. A. Garcia-Bellido and J. R. Merriam, Dev. Biol. 24, 61 (1971); M. J. Fain and B. Stevens, ibid. 92, 247 (1982); B. J. Graves and G. Schubiger, ibid. 93, 104 (1982).
30. F. C. Nielsen, L. Ostergaard, J. Nielsen, J. Christiansen, Nature 377, 358 (1995); I. B. Leibiger, B. Leibiger, T. Moede, P. O. Berggren, Mol. Cell 1, 933 (1998).
31. F. C. Nielsen, L. Ostergaard, J. Nielsen, J. Christiansen, Nature 377, 358 (1995); I. B. Leibiger, B. Leibiger, T. Moede, P. O. Berggren, Mol. Cell 1, 933 (1998).
32. Somatic clones were generated with the FLP-FRT recombination system [T. Xu and G. M. Rubin, Development 117, 1223 (1993)]. A 1-hour heat shock during the first larval instar permitted the recovery of clones in 50% of adults. Cross sections in eyes were performed as described [K. Basler, B. Christen, E. Hafen, Cell 64, 1069 (1991)].
33. Somatic clones were generated with the FLP-FRT recombination system [T. Xu and G. M. Rubin, Development 117, 1223 (1993)]. A 1-hour heat shock during the first larval instar permitted the recovery of clones in 50% of adults. Cross sections in eyes were performed as described [K. Basler, B. Christen, E. Hafen, Cell 64, 1069 (1991)].
34. A. Garcia-Bellido, P. Ripoll, G. Morata, Nature New Biol. 245, 251 (1973); D. Fristrom, M. Wilcox, J. Fristrom, Development 117, 509 (1993).
35. A. Garcia-Bellido, P. Ripoll, G. Morata, Nature New Biol. 245, 251 (1973); D. Fristrom, M. Wilcox, J. Fristrom, Development 117, 509 (1993).
36. N. Gorfinkiel, G. Morata, I. Guerrero, Genes Dev. 11, 2259 (1997); F. J. Diaz-Benjumea and S. M. Cohen, Cell 75, 741 (1993).
37. N. Gorfinkiel, G. Morata, I. Guerrero, Genes Dev. 11, 2259 (1997); F. J. Diaz-Benjumea and S. M. Cohen, Cell 75, 741 (1993).
38. A. H. Brand and N. Perrimon, Development 118, 401 (1993). Wing discs were dissected and stained with the D20 antibody (27) as described [U. G. Gaul, G. Mardon. G. M. Rubin, Cell 68, 1007 (1992)].
39. A. H. Brand and N. Perrimon, Development 118, 401 (1993). Wing discs were dissected and stained with the D20 antibody (27) as described [U. G. Gaul, G. Mardon. G. M. Rubin, Cell 68, 1007 (1992)].
40. When the wing blade is bent down such that it represents the arc of a quarter of a circle, the length of the dorsal wing sheet can be calculated from the equation 2L/π = (2L′/π) - d, where L and L′ represent the length of the ventral and dorsal wing sheets, respectively, and d is the distance between the middle of the two wing sheets. Because the length of the wing blade is 2 mm and the distance between the middle of the two wing sheets is about 10 μM [J. B. Tucker et al., Eur. J. Cell Biol. 41, 279 (1986)], the length of the dorsal sheet would measure 2.016 mm, a change of less than 1%.
41. P. B. Dennis, S. Fumagalli, G. Thomas, Curr. Opin. Genet. Dev. 9, 49 (1999).
42. S. J. Leevers, D. Weinkove, L. K. MacDougall, E. Hafen, M. D. Waterfield, EMBO J. 15, 6584 (1996).
43. M. Laser et al., J. Biol. Chem. 273, 24610 (1998).
44. M. Ashbumer, Drosophila: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989).
45. M. J. Stewart, C. O. A. Berry, F. Zilberman, G. Thomas, S. C. Kozma, Proc. Natl. Acad. Sci. U.S.A. 93, 10791 (1996).
46. The authors thank W. Gehring, M. Jacobs-Lorena, W. Krek, K. Nasmyth, and M. Raff for fruitful discussions; J. Bailey-Serres, P. Caroni, P. B. Dennis, M. Hall, N. Pullen, and S. Volarevic for critical reading of the manuscript; J. Bailey-Serres for correcting the dS6K sequence; the Bloomington Stock Center for fly strains; M. Rothnie for the preparation of figures; A. Hefti for scanning electron micrographs; and M. Dessing for FACS analysis. M.J.S. was a recipient of an NIH National Research Service Award F32 GM15926, and this work was supported in part by a grant from the Swiss Cancer League to E.H., S.C.K., and G.T.