Abstract
The Drosophila protein Nanos encodes an evolutionarily conserved protein with two zinc finger motifs. In the embryo, Nanos protein function is required for establishment of the anterior-posterior body pattern and for the migration of primordial germ cells. During oogenesis, Nanos protein is involved in the establishment and maintenance of germ-line stem cells and the differentiation of oocyte precursor cells. To establish proper embryonic patterning, Nanos acts as a translational regulator of hunchback RNA. Nanos' targets for germ cell migration and development are not known. Here, we describe a selective genetic screen aimed at isolating new nanos alleles. The molecular and genetic analysis of 68 new alleles has allowed us to identify amino acids critical for nanos function. This analysis shows that the CCHC motifs, which coordinate two metal ions, are essential for all known functions of Nanos protein. Furthermore, a region C-terminal to the zinc fingers seems to constitute a novel functional domain within the Nanos protein. This "tail region" of Nanos is required for abdomen formation and germ cell migration, but not for oogenesis.
Full Text
The Full Text of this article is available as a PDF (271.4 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahringer J., Kimble J. Control of the sperm-oocyte switch in Caenorhabditis elegans hermaphrodites by the fem-3 3' untranslated region. Nature. 1991 Jan 24;349(6307):346–348. doi: 10.1038/349346a0. [DOI] [PubMed] [Google Scholar]
- Ahringer J., Rosenquist T. A., Lawson D. N., Kimble J. The Caenorhabditis elegans sex determining gene fem-3 is regulated post-transcriptionally. EMBO J. 1992 Jun;11(6):2303–2310. doi: 10.1002/j.1460-2075.1992.tb05289.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Asaoka M., Sano H., Obara Y., Kobayashi S. Maternal Nanos regulates zygotic gene expression in germline progenitors of Drosophila melanogaster. Mech Dev. 1998 Nov;78(1-2):153–158. doi: 10.1016/s0925-4773(98)00164-6. [DOI] [PubMed] [Google Scholar]
- Bai C., Tolias P. P. Drosophila clipper/CPSF 30K is a post-transcriptionally regulated nuclear protein that binds RNA containing GC clusters. Nucleic Acids Res. 1998 Apr 1;26(7):1597–1604. doi: 10.1093/nar/26.7.1597. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barker D. D., Wang C., Moore J., Dickinson L. K., Lehmann R. Pumilio is essential for function but not for distribution of the Drosophila abdominal determinant Nanos. Genes Dev. 1992 Dec;6(12A):2312–2326. doi: 10.1101/gad.6.12a.2312. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Curtis D., Apfeld J., Lehmann R. nanos is an evolutionarily conserved organizer of anterior-posterior polarity. Development. 1995 Jun;121(6):1899–1910. doi: 10.1242/dev.121.6.1899. [DOI] [PubMed] [Google Scholar]
- Curtis D., Treiber D. K., Tao F., Zamore P. D., Williamson J. R., Lehmann R. A CCHC metal-binding domain in Nanos is essential for translational regulation. EMBO J. 1997 Feb 17;16(4):834–843. doi: 10.1093/emboj/16.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dannull J., Surovoy A., Jung G., Moelling K. Specific binding of HIV-1 nucleocapsid protein to PSI RNA in vitro requires N-terminal zinc finger and flanking basic amino acid residues. EMBO J. 1994 Apr 1;13(7):1525–1533. doi: 10.1002/j.1460-2075.1994.tb06414.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Guzman R. N., Wu Z. R., Stalling C. C., Pappalardo L., Borer P. N., Summers M. F. Structure of the HIV-1 nucleocapsid protein bound to the SL3 psi-RNA recognition element. Science. 1998 Jan 16;279(5349):384–388. doi: 10.1126/science.279.5349.384. [DOI] [PubMed] [Google Scholar]
- Ephrussi A., Dickinson L. K., Lehmann R. Oskar organizes the germ plasm and directs localization of the posterior determinant nanos. Cell. 1991 Jul 12;66(1):37–50. doi: 10.1016/0092-8674(91)90137-n. [DOI] [PubMed] [Google Scholar]
- Ephrussi A., Lehmann R. Induction of germ cell formation by oskar. Nature. 1992 Jul 30;358(6385):387–392. doi: 10.1038/358387a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Gavis E. R., Curtis D., Lehmann R. Identification of cis-acting sequences that control nanos RNA localization. Dev Biol. 1996 May 25;176(1):36–50. doi: 10.1006/dbio.1996.9996. [DOI] [PubMed] [Google Scholar]
- Gavis E. R., Lehmann R. Localization of nanos RNA controls embryonic polarity. Cell. 1992 Oct 16;71(2):301–313. doi: 10.1016/0092-8674(92)90358-j. [DOI] [PubMed] [Google Scholar]
- Gavis E. R., Lehmann R. Translational regulation of nanos by RNA localization. Nature. 1994 May 26;369(6478):315–318. doi: 10.1038/369315a0. [DOI] [PubMed] [Google Scholar]
- Gavis E. R., Lunsford L., Bergsten S. E., Lehmann R. A conserved 90 nucleotide element mediates translational repression of nanos RNA. Development. 1996 Sep;122(9):2791–2800. doi: 10.1242/dev.122.9.2791. [DOI] [PubMed] [Google Scholar]
- Hülskamp M., Schröder C., Pfeifle C., Jäckle H., Tautz D. Posterior segmentation of the Drosophila embryo in the absence of a maternal posterior organizer gene. Nature. 1989 Apr 20;338(6217):629–632. doi: 10.1038/338629a0. [DOI] [PubMed] [Google Scholar]
- Irish V., Lehmann R., Akam M. The Drosophila posterior-group gene nanos functions by repressing hunchback activity. Nature. 1989 Apr 20;338(6217):646–648. doi: 10.1038/338646a0. [DOI] [PubMed] [Google Scholar]
- Kobayashi S., Yamada M., Asaoka M., Kitamura T. Essential role of the posterior morphogen nanos for germline development in Drosophila. Nature. 1996 Apr 25;380(6576):708–711. doi: 10.1038/380708a0. [DOI] [PubMed] [Google Scholar]
- Kraemer B., Crittenden S., Gallegos M., Moulder G., Barstead R., Kimble J., Wickens M. NANOS-3 and FBF proteins physically interact to control the sperm-oocyte switch in Caenorhabditis elegans. Curr Biol. 1999 Sep 23;9(18):1009–1018. doi: 10.1016/s0960-9822(99)80449-7. [DOI] [PubMed] [Google Scholar]
- Lehmann R., Nüsslein-Volhard C. Abdominal segmentation, pole cell formation, and embryonic polarity require the localized activity of oskar, a maternal gene in Drosophila. Cell. 1986 Oct 10;47(1):141–152. doi: 10.1016/0092-8674(86)90375-2. [DOI] [PubMed] [Google Scholar]
- Lehmann R., Nüsslein-Volhard C. The maternal gene nanos has a central role in posterior pattern formation of the Drosophila embryo. Development. 1991 Jul;112(3):679–691. doi: 10.1242/dev.112.3.679. [DOI] [PubMed] [Google Scholar]
- Macdonald P. M. The Drosophila pumilio gene: an unusually long transcription unit and an unusual protein. Development. 1992 Jan;114(1):221–232. doi: 10.1242/dev.114.1.221. [DOI] [PubMed] [Google Scholar]
- Mosquera L., Forristall C., Zhou Y., King M. L. A mRNA localized to the vegetal cortex of Xenopus oocytes encodes a protein with a nanos-like zinc finger domain. Development. 1993 Jan;117(1):377–386. doi: 10.1242/dev.117.1.377. [DOI] [PubMed] [Google Scholar]
- Murata Y., Wharton R. P. Binding of pumilio to maternal hunchback mRNA is required for posterior patterning in Drosophila embryos. Cell. 1995 Mar 10;80(5):747–756. doi: 10.1016/0092-8674(95)90353-4. [DOI] [PubMed] [Google Scholar]
- Ottmann M., Gabus C., Darlix J. L. The central globular domain of the nucleocapsid protein of human immunodeficiency virus type 1 is critical for virion structure and infectivity. J Virol. 1995 Mar;69(3):1778–1784. doi: 10.1128/jvi.69.3.1778-1784.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rajavashisth T. B., Taylor A. K., Andalibi A., Svenson K. L., Lusis A. J. Identification of a zinc finger protein that binds to the sterol regulatory element. Science. 1989 Aug 11;245(4918):640–643. doi: 10.1126/science.2562787. [DOI] [PubMed] [Google Scholar]
- Smibert C. A., Wilson J. E., Kerr K., Macdonald P. M. smaug protein represses translation of unlocalized nanos mRNA in the Drosophila embryo. Genes Dev. 1996 Oct 15;10(20):2600–2609. doi: 10.1101/gad.10.20.2600. [DOI] [PubMed] [Google Scholar]
- Struhl G. Differing strategies for organizing anterior and posterior body pattern in Drosophila embryos. Nature. 1989 Apr 27;338(6218):741–744. doi: 10.1038/338741a0. [DOI] [PubMed] [Google Scholar]
- Subramaniam K., Seydoux G. nos-1 and nos-2, two genes related to Drosophila nanos, regulate primordial germ cell development and survival in Caenorhabditis elegans. Development. 1999 Nov;126(21):4861–4871. doi: 10.1242/dev.126.21.4861. [DOI] [PubMed] [Google Scholar]
- Theurkauf W. E., Hawley R. S. Meiotic spindle assembly in Drosophila females: behavior of nonexchange chromosomes and the effects of mutations in the nod kinesin-like protein. J Cell Biol. 1992 Mar;116(5):1167–1180. doi: 10.1083/jcb.116.5.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang C., Dickinson L. K., Lehmann R. Genetics of nanos localization in Drosophila. Dev Dyn. 1994 Feb;199(2):103–115. doi: 10.1002/aja.1001990204. [DOI] [PubMed] [Google Scholar]
- Wang C., Lehmann R. Nanos is the localized posterior determinant in Drosophila. Cell. 1991 Aug 23;66(4):637–647. doi: 10.1016/0092-8674(91)90110-k. [DOI] [PubMed] [Google Scholar]
- Wharton R. P., Sonoda J., Lee T., Patterson M., Murata Y. The Pumilio RNA-binding domain is also a translational regulator. Mol Cell. 1998 May;1(6):863–872. doi: 10.1016/s1097-2765(00)80085-4. [DOI] [PubMed] [Google Scholar]
- Wharton R. P., Struhl G. RNA regulatory elements mediate control of Drosophila body pattern by the posterior morphogen nanos. Cell. 1991 Nov 29;67(5):955–967. doi: 10.1016/0092-8674(91)90368-9. [DOI] [PubMed] [Google Scholar]
- Wilson R., Ainscough R., Anderson K., Baynes C., Berks M., Bonfield J., Burton J., Connell M., Copsey T., Cooper J. 2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans. Nature. 1994 Mar 3;368(6466):32–38. doi: 10.1038/368032a0. [DOI] [PubMed] [Google Scholar]
- Zamore P. D., Bartel D. P., Lehmann R., Williamson J. R. The PUMILIO-RNA interaction: a single RNA-binding domain monomer recognizes a bipartite target sequence. Biochemistry. 1999 Jan 12;38(2):596–604. doi: 10.1021/bi982264s. [DOI] [PubMed] [Google Scholar]
- Zamore P. D., Williamson J. R., Lehmann R. The Pumilio protein binds RNA through a conserved domain that defines a new class of RNA-binding proteins. RNA. 1997 Dec;3(12):1421–1433. [PMC free article] [PubMed] [Google Scholar]
- Zhang B., Gallegos M., Puoti A., Durkin E., Fields S., Kimble J., Wickens M. P. A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line. Nature. 1997 Dec 4;390(6659):477–484. doi: 10.1038/37297. [DOI] [PubMed] [Google Scholar]
