Abstract
The Caenorhabditis elegans sex-determining gene tra-2 promotes female development and expresses 4.7-, 1.9- and 1.8-kb mRNAs. The 4.7-kb mRNA encodes the major feminizing activity of the locus, a predicted membrane receptor that mediates cell-to-cell communication, named TRA-2A. The tra-2 gene was characterized from a close relative, C. briggsae. The Cb-tra-2 gene expresses only a 4.7-kb mRNA and alternatively spliced variants, which encode TRA-2A homologues. The Cb-TRA-2A and Ce-TRA-2A sequences are highly diverged, sharing only 43% identity, although their hydropathy profiles remain remarkably similar. Three potential regulatory sites of Ce-tra-2 activity were previously identified by analyzing tra-2(eg), tra-2(gf), and tra-2(mx) mutations. Two of these sites, the EG site and MX region, are conserved in Cb-tra-2. By contrast, the two direct repeat elements in the Ce-tra-2 3' untranslated region, which are disrupted in tra-2(gf) mutants, are absent. Injection of Cb-tra-2 antisense RNA into C. briggsae mimics the Ce-tra-2 loss-of-function phenotype. Thus, antisense RNA permits studies of gene activity in nematodes that lack extensive genetics.
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- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doniach T. Activity of the sex-determining gene tra-2 is modulated to allow spermatogenesis in the C. elegans hermaphrodite. Genetics. 1986 Sep;114(1):53–76. doi: 10.1093/genetics/114.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Emmons S. W., Klass M. R., Hirsh D. Analysis of the constancy of DNA sequences during development and evolution of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1333–1337. doi: 10.1073/pnas.76.3.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Goodrich L. V., Johnson R. L., Milenkovic L., McMahon J. A., Scott M. P. Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog. Genes Dev. 1996 Feb 1;10(3):301–312. doi: 10.1101/gad.10.3.301. [DOI] [PubMed] [Google Scholar]
- Goodwin E. B., Okkema P. G., Evans T. C., Kimble J. Translational regulation of tra-2 by its 3' untranslated region controls sexual identity in C. elegans. Cell. 1993 Oct 22;75(2):329–339. doi: 10.1016/0092-8674(93)80074-o. [DOI] [PubMed] [Google Scholar]
- Guo S., Kemphues K. J. par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed. Cell. 1995 May 19;81(4):611–620. doi: 10.1016/0092-8674(95)90082-9. [DOI] [PubMed] [Google Scholar]
- Hengartner M. O., Horvitz H. R. C. elegans cell survival gene ced-9 encodes a functional homolog of the mammalian proto-oncogene bcl-2. Cell. 1994 Feb 25;76(4):665–676. doi: 10.1016/0092-8674(94)90506-1. [DOI] [PubMed] [Google Scholar]
- Henikoff S. Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 1987;155:156–165. doi: 10.1016/0076-6879(87)55014-5. [DOI] [PubMed] [Google Scholar]
- Heschl M. F., Baillie D. L. Functional elements and domains inferred from sequence comparisons of a heat shock gene in two nematodes. J Mol Evol. 1990 Jul;31(1):3–9. doi: 10.1007/BF02101786. [DOI] [PubMed] [Google Scholar]
- Hinnebusch A. G. Gene-specific translational control of the yeast GCN4 gene by phosphorylation of eukaryotic initiation factor 2. Mol Microbiol. 1993 Oct;10(2):215–223. doi: 10.1111/j.1365-2958.1993.tb01947.x. [DOI] [PubMed] [Google Scholar]
- Hodgkin J. A genetic analysis of the sex-determining gene, tra-1, in the nematode Caenorhabditis elegans. Genes Dev. 1987 Sep;1(7):731–745. doi: 10.1101/gad.1.7.731. [DOI] [PubMed] [Google Scholar]
- Hodgkin J., Albertson D. G. Isolation of dominant XO-feminizing mutations in Caenorhabditis elegans: new regulatory tra alleles and an X chromosome duplication with implications for primary sex determination. Genetics. 1995 Oct;141(2):527–542. doi: 10.1093/genetics/141.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hodgkin J. Genetic nomenclature guide. Caenorhabditis elegans. Trends Genet. 1995 Mar;:24–25. [PubMed] [Google Scholar]
- Hodgkin J. Molecular cloning and duplication of the nematode sex-determining gene tra-1. Genetics. 1993 Mar;133(3):543–560. doi: 10.1093/genetics/133.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hodgkin J. Sex determination and dosage compensation in Caenorhabditis elegans. Annu Rev Genet. 1987;21:133–154. doi: 10.1146/annurev.ge.21.120187.001025. [DOI] [PubMed] [Google Scholar]
- Hodgkin J. Sex determination compared in Drosophila and Caenorhabditis. Nature. 1990 Apr 19;344(6268):721–728. doi: 10.1038/344721a0. [DOI] [PubMed] [Google Scholar]
- Huang X. Y., Hirsh D. A second trans-spliced RNA leader sequence in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8640–8644. doi: 10.1073/pnas.86.22.8640. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunter C. P., Wood W. B. Evidence from mosaic analysis of the masculinizing gene her-1 for cell interactions in C. elegans sex determination. Nature. 1992 Feb 6;355(6360):551–555. doi: 10.1038/355551a0. [DOI] [PubMed] [Google Scholar]
- Koopman P., Gubbay J., Vivian N., Goodfellow P., Lovell-Badge R. Male development of chromosomally female mice transgenic for Sry. Nature. 1991 May 9;351(6322):117–121. doi: 10.1038/351117a0. [DOI] [PubMed] [Google Scholar]
- Kuwabara P. E. A novel regulatory mutation in the C. elegans sex determination gene tra-2 defines a candidate ligand/receptor interaction site. Development. 1996 Jul;122(7):2089–2098. doi: 10.1242/dev.122.7.2089. [DOI] [PubMed] [Google Scholar]
- Kuwabara P. E., Kimble J. A predicted membrane protein, TRA-2A, directs hermaphrodite development in Caenorhabditis elegans. Development. 1995 Sep;121(9):2995–3004. doi: 10.1242/dev.121.9.2995. [DOI] [PubMed] [Google Scholar]
- Kuwabara P. E., Kimble J. Molecular genetics of sex determination in C. elegans. Trends Genet. 1992 May;8(5):164–168. doi: 10.1016/0168-9525(92)90218-s. [DOI] [PubMed] [Google Scholar]
- Kuwabara P. E., Okkema P. G., Kimble J. tra-2 encodes a membrane protein and may mediate cell communication in the Caenorhabditis elegans sex determination pathway. Mol Biol Cell. 1992 Apr;3(4):461–473. doi: 10.1091/mbc.3.4.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuwabara P. E., Shah S. Cloning by synteny: identifying C. briggsae homologues of C. elegans genes. Nucleic Acids Res. 1994 Oct 25;22(21):4414–4418. doi: 10.1093/nar/22.21.4414. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee Y. H., Huang X. Y., Hirsh D., Fox G. E., Hecht R. M. Conservation of gene organization and trans-splicing in the glyceraldehyde-3-phosphate dehydrogenase-encoding genes of Caenorhabditis briggsae. Gene. 1992 Nov 16;121(2):227–235. doi: 10.1016/0378-1119(92)90126-a. [DOI] [PubMed] [Google Scholar]
- Mello C. C., Kramer J. M., Stinchcomb D., Ambros V. Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences. EMBO J. 1991 Dec;10(12):3959–3970. doi: 10.1002/j.1460-2075.1991.tb04966.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okkema P. G., Kimble J. Molecular analysis of tra-2, a sex determining gene in C.elegans. EMBO J. 1991 Jan;10(1):171–176. doi: 10.1002/j.1460-2075.1991.tb07933.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perry M. D., Li W., Trent C., Robertson B., Fire A., Hageman J. M., Wood W. B. Molecular characterization of the her-1 gene suggests a direct role in cell signaling during Caenorhabditis elegans sex determination. Genes Dev. 1993 Feb;7(2):216–228. doi: 10.1101/gad.7.2.216. [DOI] [PubMed] [Google Scholar]
- Sinclair A. H., Berta P., Palmer M. S., Hawkins J. R., Griffiths B. L., Smith M. J., Foster J. W., Frischauf A. M., Lovell-Badge R., Goodfellow P. N. A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif. Nature. 1990 Jul 19;346(6281):240–244. doi: 10.1038/346240a0. [DOI] [PubMed] [Google Scholar]
- Spieth J., Brooke G., Kuersten S., Lea K., Blumenthal T. Operons in C. elegans: polycistronic mRNA precursors are processed by trans-splicing of SL2 to downstream coding regions. Cell. 1993 May 7;73(3):521–532. doi: 10.1016/0092-8674(93)90139-h. [DOI] [PubMed] [Google Scholar]
- Villeneuve A. M., Meyer B. J. The regulatory hierarchy controlling sex determination and dosage compensation in Caenorhabditis elegans. Adv Genet. 1990;27:117–188. doi: 10.1016/s0065-2660(08)60025-5. [DOI] [PubMed] [Google Scholar]
- Zarkower D., Hodgkin J. Molecular analysis of the C. elegans sex-determining gene tra-1: a gene encoding two zinc finger proteins. Cell. 1992 Jul 24;70(2):237–249. doi: 10.1016/0092-8674(92)90099-x. [DOI] [PubMed] [Google Scholar]
- de Bono M., Hodgkin J. Evolution of sex determination in caenorhabditis: unusually high divergence of tra-1 and its functional consequences. Genetics. 1996 Oct;144(2):587–595. doi: 10.1093/genetics/144.2.587. [DOI] [PMC free article] [PubMed] [Google Scholar]