Still life, a protein in synaptic terminals of Drosophila homologous to GDP-GTP exchangers

Science

Volumn 275, Issue 5299, 1997, Pages 543-547

  Sone, Masaki ^a   Hoshino, Mikio ^a   Suzuki, Emiko ^a   Kuroda, Shinya ^a   Kaibuchi, Kozo ^a   Nakagoshi, Hideki ^a   Saigo, Kaoru ^a   Nabeshima, Yo Ichi ^a   Hama, Chihiro ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

PROTEIN;

ARTICLE; DROSOPHILA; NERVE ENDING; NONHUMAN; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN EXPRESSION; SYNAPSE;

ACTINS; AMINO ACID SEQUENCE; ANIMALS; AXONS; CELL MEMBRANE; CYTOSKELETON; DNA, COMPLEMENTARY; DROSOPHILA; DROSOPHILA PROTEINS; EMBRYO, NONMAMMALIAN; GENE EXPRESSION; GENES, INSECT; GTP PHOSPHOHYDROLASES; GTP-BINDING PROTEINS; GUANINE NUCLEOTIDE EXCHANGE FACTORS; HUMANS; IN SITU HYBRIDIZATION; KB CELLS; MOLECULAR SEQUENCE DATA; MOVEMENT; MUTATION; NEUROMUSCULAR JUNCTION; PRESYNAPTIC TERMINALS; RAC GTP-BINDING PROTEINS; SIGNAL TRANSDUCTION;

EID: 0031037406     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5299.543     Document Type: Article

References (42)

1. Z. W. Hall and J. R. Sanes, Cell/Neuron 72/10 (suppl.), 99 (1993); M. E. Burns and G. J. Augustine, Cell 83, 187 (1995); R. J. Kleiman and L. F. Reichardt, ibid. 85, 461 (1996); D. C. Lo, Neuron 15, 979 (1995); C. S. Goodman and C. J. Shatz, Cell/ Neuron 72/10 (suppl.), 77 (1993).
2. Z. W. Hall and J. R. Sanes, Cell/Neuron 72/10 (suppl.), 99 (1993); M. E. Burns and G. J. Augustine, Cell 83, 187 (1995); R. J. Kleiman and L. F. Reichardt, ibid. 85, 461 (1996); D. C. Lo, Neuron 15, 979 (1995); C. S. Goodman and C. J. Shatz, Cell/ Neuron 72/10 (suppl.), 77 (1993).
3. Z. W. Hall and J. R. Sanes, Cell/Neuron 72/10 (suppl.), 99 (1993); M. E. Burns and G. J. Augustine, Cell 83, 187 (1995); R. J. Kleiman and L. F. Reichardt, ibid. 85, 461 (1996); D. C. Lo, Neuron 15, 979 (1995); C. S. Goodman and C. J. Shatz, Cell/ Neuron 72/10 (suppl.), 77 (1993).
4. Z. W. Hall and J. R. Sanes, Cell/Neuron 72/10 (suppl.), 99 (1993); M. E. Burns and G. J. Augustine, Cell 83, 187 (1995); R. J. Kleiman and L. F. Reichardt, ibid. 85, 461 (1996); D. C. Lo, Neuron 15, 979 (1995); C. S. Goodman and C. J. Shatz, Cell/ Neuron 72/10 (suppl.), 77 (1993).
5. Z. W. Hall and J. R. Sanes, Cell/Neuron 72/10 (suppl.), 99 (1993); M. E. Burns and G. J. Augustine, Cell 83, 187 (1995); R. J. Kleiman and L. F. Reichardt, ibid. 85, 461 (1996); D. C. Lo, Neuron 15, 979 (1995); C. S. Goodman and C. J. Shatz, Cell/ Neuron 72/10 (suppl.), 77 (1993).
6. A. Hall, Annu. Rev. Cell Biol. 10, 31 (1994); L. M. Machesky and A. Hall, Trends Cell Biol. 6, 304 (1996).
7. A. Hall, Annu. Rev. Cell Biol. 10, 31 (1994); L. M. Machesky and A. Hall, Trends Cell Biol. 6, 304 (1996).
8. L. Luo et al., Nature 379, 837 (1996); D. J. G. Mackay, C. D. Nobes, A. Hall, Trends Neurosci. 18, 496 (1995).
9. L. Luo et al., Nature 379, 837 (1996); D. J. G. Mackay, C. D. Nobes, A. Hall, Trends Neurosci. 18, 496 (1995).
10. L. Luo, Y. J. Liao, L.Y. Jan, Y. N. Jan, Genes Dev. 8, 1787 (1994).
11. M. Hoshino, F. Matsuzaki, Y.-i. Nabeshima, C. Hama, Neuron 10, 395 (1993).
12. M. Hoshino, E. Suzuki, Y.-i. Nabeshima, C. Hama, Development 122, 589 (1996).
13. 98.1 mutant was constructed by insertion of the type 1 sif cDNA into a transformation vector containing the 3.5-kb promoter fragment of the elav gene [K.-M. Yao and K. White, J. Neurochem. 63, 41 (1994)] and the hsp70 polyadenylate additional signal. This plasmid was introduced to white flies by P element-mediated germline transformation.
14. The nucleotide sequence data reported here will appear in the DNA Databank of Japan, European Molecular Biology Laboratory, and GenBank with the accession numbers D86546 and D86547. In both cDNAs, the nucleotide sequences around the first ATG codon match the Drosophila translation start consensus sequence [D. R. Cavener, Nucleic Acids Res. 15, 1353 (1987)]. In the 2064-amino acid protein, an in-frame stop codon is located 54 bp upstream of the ATG codon. In the 2044-amino acid protein, the location of an in-frame stop codon 945 bp upstream of the ATG codon raises the possibility that a codon other than ATG is used as the initiation codon as previously reported for several genes [M. B. Feany and W. G. Quinn, Science 268, 869 (1995)].
15. The nucleotide sequence data reported here will appear in the DNA Databank of Japan, European Molecular Biology Laboratory, and GenBank with the accession numbers D86546 and D86547. In both cDNAs, the nucleotide sequences around the first ATG codon match the Drosophila translation start consensus sequence [D. R. Cavener, Nucleic Acids Res. 15, 1353 (1987)]. In the 2064-amino acid protein, an in-frame stop codon is located 54 bp upstream of the ATG codon. In the 2044-amino acid protein, the location of an in-frame stop codon 945 bp upstream of the ATG codon raises the possibility that a codon other than ATG is used as the initiation codon as previously reported for several genes [M. B. Feany and W. G. Quinn, Science 268, 869 (1995)].
16. G. G. M. Habets et al., Cell 77, 537 (1994).
17. M. A. Lemmon, K. M. Ferguson, J. Schlessinger, ibid. 85, 621 (1996).
18. M. Rechsteiner and S. W. Rogers, Trends Biochem. Sci. 21, 267 (1996).
19. C. P. Ponting and C. Phillips, ibid. 20, 102 (1995); J. E. Brenman et al., Cell 84, 757 (1996).
20. C. P. Ponting and C. Phillips, ibid. 20, 102 (1995); J. E. Brenman et al., Cell 84, 757 (1996).
21. M. J. Hart et al., J. Biol. Chem. 269, 62 (1994); L. A. Quilliam, R. Khosravi-Far, S. Y. Huff, C. J. Der, BioEssays 17, 395 (1995).
22. M. J. Hart et al., J. Biol. Chem. 269, 62 (1994); L. A. Quilliam, R. Khosravi-Far, S. Y. Huff, C. J. Der, BioEssays 17, 395 (1995).
23. F. Michiels, G. G. M. Habets, J. C. Stam, R. A. van der Kammen, J. G. Collard, Nature 375, 338 (1995).
24. In situ hybridization to embryos was performed as previously described (5) with a single-stranded DNA probe corresponding to amino acids 870 to 1095. Embryonic stages are as previously described [J. A. Campos-Ortega and V. Hartenstein, The Embryonic Development of Drosophila melanogaster (Springer-Verlag, Berlin, 1985)].
25. J. T. Littleton, H. J. Bellen, M. S. Perin, Development 118, 1077 (1993).
26. Two antibodies to SIF, AbI3 and AbH3, were raised to fusion proteins that consisted of portions of the SIF proteins (amino acids 327 to 604 of type 2 for AbI3 and 1080 to 1269 for AbH3) and metal-binding domains derived from pET16b (Novagen) and pTrcHis A (Invitrogen) vector, respectively. Injection of the purified proteins into rats and affinity purification of the antisera were performed as described (6). For immunostaining, embryos and larvae were fixed with 4% paraformaldehyde in phosphate-buffered saline for 20 min and stained with antibody to SIF using ABC Elite (Vector Labs). Preparation and staining of adult brain sections and immunoelectron microscopy were performed as described (6).
27. In embryos and larvae, we observed no staining with AbH3 probably because of insufficient SIF expression for the sensitivity of this antibody.
28. I. A. Meinertzhagen and S. D. O'Neil, J. Comp. Neurol. 305, 232 (1991).
29. The GAL4-UAS system was used to express the transgene in the embryonic and larval nervous systems [A. H. Brand and N. Perrimon, Development 118, 401 (1993)].
30. The plasmid expressing SIFΔN protein was constructed by ligating cDNA fragments encoding a portion of the kinesin heavy chain (amino acids 1 to 605) [E. Giniger, W. Wells, L. Y. Jan, Y. N. Jan, Roux's Arch. Dev. Biol. 202, 112 (1993); M. F. A. VanBerkum and C. S. Goodman, Neuron 14, 43 (1995)] and a portion of SIF (amino acids 838 to 2064) into the pUAST transforming vector (20).
31. The plasmid expressing SIFΔN protein was constructed by ligating cDNA fragments encoding a portion of the kinesin heavy chain (amino acids 1 to 605) [E. Giniger, W. Wells, L. Y. Jan, Y. N. Jan, Roux's Arch. Dev. Biol. 202, 112 (1993); M. F. A. VanBerkum and C. S. Goodman, Neuron 14, 43 (1995)] and a portion of SIF (amino acids 838 to 2064) into the pUAST transforming vector (20).
32. D. Van Vactor, H. Sink, D. Fambrough, R. Tsoo, C. S. Goodman, Cell 73, 1137 (1993).
33. This strain dies during the embryonic and first instar larval stages. Two other independent embryonic lethal lines carrying S/FΔN exhibit similar but more severe phenotypes.
34. J. Johansen, M. E. Halpern, K. M. Johansen, H. Keshishian, J. Neurosci. 9, 710 (1989).
35. M. Sone et al., data not shown; T. Nishiyama et al., Mol. Cell. Biol. 14, 2447 (1994); C. D. Nobes and A. Hall, Cell 81, 53 (1995).
36. M. Sone et al., data not shown; T. Nishiyama et al., Mol. Cell. Biol. 14, 2447 (1994); C. D. Nobes and A. Hall, Cell 81, 53 (1995).
37. M. Sone et al., data not shown; T. Nishiyama et al., Mol. Cell. Biol. 14, 2447 (1994); C. D. Nobes and A. Hall, Cell 81, 53 (1995).
38. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
39. The PEST scores are 8.88 (type 1), 6.62 (type 2, amino acids 389 to 409), and 9.18 (type 2, amino acids 534 to 563).
40. L. Y. Jan and Y. N. Jan, Proc. Natl. Acad. Sci. U.S.A. 79, 2700 (1982).
41. Microinjection of plasmid DNAs into the nuclei of KB cells was performed as previously described except that the cells were serum-starved for 30 hours [S. Kuroda et al., J. Biol. Chem. 271, 23363 (1996)]. The plasmid DNA, pFCMVsifΔN, used for microinjection was constructed by inserting a DNA fragment encoding portions of SIF (amino acids 1 to 18 and 838 to 2064) into pFLAG-CMV-2 (Kodak). The FLAG-SIF fusion protein was labeled with antibody to FLAG M2 (Kodak) and detected with fluorescein isothiocyanate (FITC)-conjugated antibody to mouse immunoglobulin G.
42. We thank L. Luo and Y. N. Jan for the transgenic strains carrying Drac1 and Dcdc42 derivatives, T. Uemura for the elav-GAL4 strain, A. Nose for monoclonal antibody 1D4, A. DiAntonio and E. Giniger for the plasmids containing the elav promoter and kinesin gene, respectively, C.-S. Yoon for technical instruction, and S. Yoshida, A. Chiba, and all members of our department for comments and discussions. Supported by a Grant-in-Aid for Priority Area from the Ministry of Education, Science, Sports and Culture and the research grant for Nervous and Mental Disorders from the Ministry of Health and Welfare of Japan.