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. 2000 Sep;156(1):245–256. doi: 10.1093/genetics/156.1.245

The shut-down gene of Drosophila melanogaster encodes a novel FK506-binding protein essential for the formation of germline cysts during oogenesis.

K Munn 1, R Steward 1
PMCID: PMC1461232  PMID: 10978289

Abstract

In Drosophila melanogaster, the process of oogenesis is initiated with the asymmetric division of a germline stem cell. This division results in the self-renewal of the stem cell and the generation of a daughter cell that undergoes four successive mitotic divisions to produce a germline cyst of 16 cells. Here, we show that shut-down is essential for the normal function of the germline stem cells. Analysis of weak loss-of-function alleles confirms that shut-down is also required at later stages of oogenesis. Clonal analysis indicates that shut-down functions autonomously in the germline. Using a positional cloning approach, we have isolated the shut-down gene. Consistent with its function, the RNA and protein are strongly expressed in the germline stem cells and in 16-cell cysts. The RNA is also present in the germ cells throughout embryogenesis. shut-down encodes a novel Drosophila protein similar to the heat-shock protein-binding immunophilins. Like immunophilins, Shut-down contains an FK506-binding protein domain and a tetratricopeptide repeat. In plants, high-molecular-weight immunophilins have been shown to regulate cell divisions in the root meristem in response to extracellular signals. Our results suggest that shut-down may regulate germ cell divisions in the germarium.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. doi: 10.1093/nar/25.17.3389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bhat K. M. The posterior determinant gene nanos is required for the maintenance of the adult germline stem cells during Drosophila oogenesis. Genetics. 1999 Apr;151(4):1479–1492. doi: 10.1093/genetics/151.4.1479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bopp D., Horabin J. I., Lersch R. A., Cline T. W., Schedl P. Expression of the Sex-lethal gene is controlled at multiple levels during Drosophila oogenesis. Development. 1993 Jul;118(3):797–812. doi: 10.1242/dev.118.3.797. [DOI] [PubMed] [Google Scholar]
  4. Brown E. J., Schreiber S. L. A signaling pathway to translational control. Cell. 1996 Aug 23;86(4):517–520. doi: 10.1016/s0092-8674(00)80125-7. [DOI] [PubMed] [Google Scholar]
  5. Cavener D. R. Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res. 1987 Feb 25;15(4):1353–1361. doi: 10.1093/nar/15.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chou T. B., Perrimon N. The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster. Genetics. 1996 Dec;144(4):1673–1679. doi: 10.1093/genetics/144.4.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chou T. B., Perrimon N. Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila. Genetics. 1992 Jul;131(3):643–653. doi: 10.1093/genetics/131.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cox D. N., Chao A., Baker J., Chang L., Qiao D., Lin H. A novel class of evolutionarily conserved genes defined by piwi are essential for stem cell self-renewal. Genes Dev. 1998 Dec 1;12(23):3715–3727. doi: 10.1101/gad.12.23.3715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ding D., Parkhurst S. M., Halsell S. R., Lipshitz H. D. Dynamic Hsp83 RNA localization during Drosophila oogenesis and embryogenesis. Mol Cell Biol. 1993 Jun;13(6):3773–3781. doi: 10.1128/mcb.13.6.3773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ewart A. K., Morris C. A., Atkinson D., Jin W., Sternes K., Spallone P., Stock A. D., Leppert M., Keating M. T. Hemizygosity at the elastin locus in a developmental disorder, Williams syndrome. Nat Genet. 1993 Sep;5(1):11–16. doi: 10.1038/ng0993-11. [DOI] [PubMed] [Google Scholar]
  11. Faure J. D., Vittorioso P., Santoni V., Fraisier V., Prinsen E., Barlier I., Van Onckelen H., Caboche M., Bellini C. The PASTICCINO genes of Arabidopsis thaliana are involved in the control of cell division and differentiation. Development. 1998 Mar;125(5):909–918. doi: 10.1242/dev.125.5.909. [DOI] [PubMed] [Google Scholar]
  12. Fischer G., Schmid F. X. The mechanism of protein folding. Implications of in vitro refolding models for de novo protein folding and translocation in the cell. Biochemistry. 1990 Mar 6;29(9):2205–2212. doi: 10.1021/bi00461a001. [DOI] [PubMed] [Google Scholar]
  13. Forbes A., Lehmann R. Nanos and Pumilio have critical roles in the development and function of Drosophila germline stem cells. Development. 1998 Feb;125(4):679–690. doi: 10.1242/dev.125.4.679. [DOI] [PubMed] [Google Scholar]
  14. Gateff E. Gonial cell neoplasm of genetic origin affecting both sexes of drosophila melanogaster. Prog Clin Biol Res. 1982;85(Pt B):621–632. [PubMed] [Google Scholar]
  15. Hofmann K., Bucher P., Falquet L., Bairoch A. The PROSITE database, its status in 1999. Nucleic Acids Res. 1999 Jan 1;27(1):215–219. doi: 10.1093/nar/27.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kay J. E. Structure-function relationships in the FK506-binding protein (FKBP) family of peptidylprolyl cis-trans isomerases. Biochem J. 1996 Mar 1;314(Pt 2):361–385. [PMC free article] [PubMed] [Google Scholar]
  17. King F. J., Lin H. Somatic signaling mediated by fs(1)Yb is essential for germline stem cell maintenance during Drosophila oogenesis. Development. 1999 May;126(9):1833–1844. doi: 10.1242/dev.126.9.1833. [DOI] [PubMed] [Google Scholar]
  18. Lamb J. R., Tugendreich S., Hieter P. Tetratrico peptide repeat interactions: to TPR or not to TPR? Trends Biochem Sci. 1995 Jul;20(7):257–259. doi: 10.1016/s0968-0004(00)89037-4. [DOI] [PubMed] [Google Scholar]
  19. Lantz V., Chang J. S., Horabin J. I., Bopp D., Schedl P. The Drosophila orb RNA-binding protein is required for the formation of the egg chamber and establishment of polarity. Genes Dev. 1994 Mar 1;8(5):598–613. doi: 10.1101/gad.8.5.598. [DOI] [PubMed] [Google Scholar]
  20. Lasko P. F., Ashburner M. Posterior localization of vasa protein correlates with, but is not sufficient for, pole cell development. Genes Dev. 1990 Jun;4(6):905–921. doi: 10.1101/gad.4.6.905. [DOI] [PubMed] [Google Scholar]
  21. Lavoie C. A., Ohlstein B., McKearin D. M. Localization and function of Bam protein require the benign gonial cell neoplasm gene product. Dev Biol. 1999 Aug 15;212(2):405–413. doi: 10.1006/dbio.1999.9346. [DOI] [PubMed] [Google Scholar]
  22. Lin H., Spradling A. C. A novel group of pumilio mutations affects the asymmetric division of germline stem cells in the Drosophila ovary. Development. 1997 Jun;124(12):2463–2476. doi: 10.1242/dev.124.12.2463. [DOI] [PubMed] [Google Scholar]
  23. Lin H. The tao of stem cells in the germline. Annu Rev Genet. 1997;31:455–491. doi: 10.1146/annurev.genet.31.1.455. [DOI] [PubMed] [Google Scholar]
  24. Lin H., Yue L., Spradling A. C. The Drosophila fusome, a germline-specific organelle, contains membrane skeletal proteins and functions in cyst formation. Development. 1994 Apr;120(4):947–956. doi: 10.1242/dev.120.4.947. [DOI] [PubMed] [Google Scholar]
  25. Mach J. M., Lehmann R. An Egalitarian-BicaudalD complex is essential for oocyte specification and axis determination in Drosophila. Genes Dev. 1997 Feb 15;11(4):423–435. doi: 10.1101/gad.11.4.423. [DOI] [PubMed] [Google Scholar]
  26. McKearin D. M., Spradling A. C. bag-of-marbles: a Drosophila gene required to initiate both male and female gametogenesis. Genes Dev. 1990 Dec;4(12B):2242–2251. doi: 10.1101/gad.4.12b.2242. [DOI] [PubMed] [Google Scholar]
  27. McKearin D., Ohlstein B. A role for the Drosophila bag-of-marbles protein in the differentiation of cystoblasts from germline stem cells. Development. 1995 Sep;121(9):2937–2947. doi: 10.1242/dev.121.9.2937. [DOI] [PubMed] [Google Scholar]
  28. Meng X., Lu X., Morris C. A., Keating M. T. A novel human gene FKBP6 is deleted in Williams syndrome. Genomics. 1998 Sep 1;52(2):130–137. doi: 10.1006/geno.1998.5412. [DOI] [PubMed] [Google Scholar]
  29. Owens-Grillo J. K., Czar M. J., Hutchison K. A., Hoffmann K., Perdew G. H., Pratt W. B. A model of protein targeting mediated by immunophilins and other proteins that bind to hsp90 via tetratricopeptide repeat domains. J Biol Chem. 1996 Jun 7;271(23):13468–13475. doi: 10.1074/jbc.271.23.13468. [DOI] [PubMed] [Google Scholar]
  30. Peattie D. A., Harding M. W., Fleming M. A., DeCenzo M. T., Lippke J. A., Livingston D. J., Benasutti M. Expression and characterization of human FKBP52, an immunophilin that associates with the 90-kDa heat shock protein and is a component of steroid receptor complexes. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10974–10978. doi: 10.1073/pnas.89.22.10974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perrot-Applanat M., Cibert C., Géraud G., Renoir J. M., Baulieu E. E. The 59 kDa FK506-binding protein, a 90 kDa heat shock protein binding immunophilin (FKBP59-HBI), is associated with the nucleus, the cytoskeleton and mitotic apparatus. J Cell Sci. 1995 May;108(Pt 5):2037–2051. doi: 10.1242/jcs.108.5.2037. [DOI] [PubMed] [Google Scholar]
  32. Pratt W. B., Czar M. J., Stancato L. F., Owens J. K. The hsp56 immunophilin component of steroid receptor heterocomplexes: could this be the elusive nuclear localization signal-binding protein? J Steroid Biochem Mol Biol. 1993 Sep;46(3):269–279. doi: 10.1016/0960-0760(93)90216-j. [DOI] [PubMed] [Google Scholar]
  33. Robinson D. N., Cooley L. Examination of the function of two kelch proteins generated by stop codon suppression. Development. 1997 Apr;124(7):1405–1417. doi: 10.1242/dev.124.7.1405. [DOI] [PubMed] [Google Scholar]
  34. Rongo C., Lehmann R. Regulated synthesis, transport and assembly of the Drosophila germ plasm. Trends Genet. 1996 Mar;12(3):102–109. doi: 10.1016/0168-9525(96)81421-1. [DOI] [PubMed] [Google Scholar]
  35. Sanchez E. R., Faber L. E., Henzel W. J., Pratt W. B. The 56-59-kilodalton protein identified in untransformed steroid receptor complexes is a unique protein that exists in cytosol in a complex with both the 70- and 90-kilodalton heat shock proteins. Biochemistry. 1990 May 29;29(21):5145–5152. doi: 10.1021/bi00473a021. [DOI] [PubMed] [Google Scholar]
  36. Schüpbach T., Wieschaus E. Female sterile mutations on the second chromosome of Drosophila melanogaster. I. Maternal effect mutations. Genetics. 1989 Jan;121(1):101–117. doi: 10.1093/genetics/121.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schüpbach T., Wieschaus E. Female sterile mutations on the second chromosome of Drosophila melanogaster. II. Mutations blocking oogenesis or altering egg morphology. Genetics. 1991 Dec;129(4):1119–1136. doi: 10.1093/genetics/129.4.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Silverstein A. M., Galigniana M. D., Kanelakis K. C., Radanyi C., Renoir J. M., Pratt W. B. Different regions of the immunophilin FKBP52 determine its association with the glucocorticoid receptor, hsp90, and cytoplasmic dynein. J Biol Chem. 1999 Dec 24;274(52):36980–36986. doi: 10.1074/jbc.274.52.36980. [DOI] [PubMed] [Google Scholar]
  39. Spradling A. C., Rubin G. M. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982 Oct 22;218(4570):341–347. doi: 10.1126/science.6289435. [DOI] [PubMed] [Google Scholar]
  40. Styhler S., Nakamura A., Swan A., Suter B., Lasko P. vasa is required for GURKEN accumulation in the oocyte, and is involved in oocyte differentiation and germline cyst development. Development. 1998 May;125(9):1569–1578. doi: 10.1242/dev.125.9.1569. [DOI] [PubMed] [Google Scholar]
  41. Suter B., Romberg L. M., Steward R. Bicaudal-D, a Drosophila gene involved in developmental asymmetry: localized transcript accumulation in ovaries and sequence similarity to myosin heavy chain tail domains. Genes Dev. 1989 Dec;3(12A):1957–1968. doi: 10.1101/gad.3.12a.1957. [DOI] [PubMed] [Google Scholar]
  42. Suter B., Steward R. Requirement for phosphorylation and localization of the Bicaudal-D protein in Drosophila oocyte differentiation. Cell. 1991 Nov 29;67(5):917–926. doi: 10.1016/0092-8674(91)90365-6. [DOI] [PubMed] [Google Scholar]
  43. Tautz D., Pfeifle C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma. 1989 Aug;98(2):81–85. doi: 10.1007/BF00291041. [DOI] [PubMed] [Google Scholar]
  44. Theurkauf W. E., Alberts B. M., Jan Y. N., Jongens T. A. A central role for microtubules in the differentiation of Drosophila oocytes. Development. 1993 Aug;118(4):1169–1180. doi: 10.1242/dev.118.4.1169. [DOI] [PubMed] [Google Scholar]
  45. Tirronen M., Partanen M., Heino T. O., Heino T. I., Roos C. Analyses of the Drosophila quit, ovarian tumor and shut down mutants in oocyte differentiation using in situ hybridisation. Mech Dev. 1993 Jan;40(1-2):113–126. doi: 10.1016/0925-4773(93)90092-c. [DOI] [PubMed] [Google Scholar]
  46. Tomancak P., Guichet A., Zavorszky P., Ephrussi A. Oocyte polarity depends on regulation of gurken by Vasa. Development. 1998 May;125(9):1723–1732. doi: 10.1242/dev.125.9.1723. [DOI] [PubMed] [Google Scholar]
  47. Vittorioso P., Cowling R., Faure J. D., Caboche M., Bellini C. Mutation in the Arabidopsis PASTICCINO1 gene, which encodes a new FK506-binding protein-like protein, has a dramatic effect on plant development. Mol Cell Biol. 1998 May;18(5):3034–3043. doi: 10.1128/mcb.18.5.3034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Xie T., Spradling A. C. decapentaplegic is essential for the maintenance and division of germline stem cells in the Drosophila ovary. Cell. 1998 Jul 24;94(2):251–260. doi: 10.1016/s0092-8674(00)81424-5. [DOI] [PubMed] [Google Scholar]
  49. Yue L., Spradling A. C. hu-li tai shao, a gene required for ring canal formation during Drosophila oogenesis, encodes a homolog of adducin. Genes Dev. 1992 Dec;6(12B):2443–2454. doi: 10.1101/gad.6.12b.2443. [DOI] [PubMed] [Google Scholar]
  50. Zaccai M., Lipshitz H. D. Differential distributions of two adducin-like protein isoforms in the Drosophila ovary and early embryo. Zygote. 1996 May;4(2):159–166. doi: 10.1017/s096719940000304x. [DOI] [PubMed] [Google Scholar]
  51. de Cuevas M., Lilly M. A., Spradling A. C. Germline cyst formation in Drosophila. Annu Rev Genet. 1997;31:405–428. doi: 10.1146/annurev.genet.31.1.405. [DOI] [PubMed] [Google Scholar]

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