Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1995 Nov;141(3):1101–1111. doi: 10.1093/genetics/141.3.1101

Two Distinct Temperature-Sensitive Alleles at the Elav Locus of Drosophila Are Suppressed Nonsense Mutations of the Same Tryptophan Codon

M L Samson 1, M J Lisbin 1, K White 1
PMCID: PMC1206833  PMID: 8582616

Abstract

The Drosophila gene elav encodes a 483-amino-acid-long nuclear RNA binding protein required for normal neuronal differentiation and maintenance. We molecularly analyzed the three known viable alleles of the gene, namely elav(ts1), elav(FliJ1), and elav(FliJ2), which manifest temperature-sensitive phenotypes. The modification of the elav(FliJ1) allele corresponds to the change of glycine(426) (GGA) into a glutamic acid (GAA). Surprisingly, elav(ts1) and elav(FliJ2) were both found to have tryptophan(419) (TGG) changed into two different stop codons, TAG and TGA, respectively. Unexpectedly, protein analysis from elav(ts1) and elav(FliJ2) reveals not only the predicted 45-kD truncated ELAV protein due to translational truncation, but also a predominant full-size 50-kD ELAV protein, both at permissive and nonpermissive temperatures. The full-length protein present in elav(ts1) and elav(FliJ2) can a priori be explained by one of several mechanisms leading to functional suppression of the nonsense mutation or by detection of a previously unrecognized ELAV isoform of similar size resulting from alternative splicing and unaffected by the stop codon. Experiments described in this article support the functional suppression of the nonsense mutation as the mechanism responsible for the full-length protein.

Full Text

The Full Text of this article is available as a PDF (5.3 MB).

Selected References

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

  1. Bandziulis R. J., Swanson M. S., Dreyfuss G. RNA-binding proteins as developmental regulators. Genes Dev. 1989 Apr;3(4):431–437. doi: 10.1101/gad.3.4.431. [DOI] [PubMed] [Google Scholar]
  2. Bentley R. C., Keene J. D. Recognition of U1 and U2 small nuclear RNAs can be altered by a 5-amino-acid segment in the U2 small nuclear ribonucleoprotein particle (snRNP) B" protein and through interactions with U2 snRNP-A' protein. Mol Cell Biol. 1991 Apr;11(4):1829–1839. doi: 10.1128/mcb.11.4.1829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bingham P. M., Chapman C. H. Evidence that white-blood is a novel type of temperature-sensitive mutation resulting from temperature-dependent effects of a transposon insertion on formation of white transcripts. EMBO J. 1986 Dec 1;5(12):3343–3351. doi: 10.1002/j.1460-2075.1986.tb04649.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Campos A. R., Grossman D., White K. Mutant alleles at the locus elav in Drosophila melanogaster lead to nervous system defects. A developmental-genetic analysis. J Neurogenet. 1985 Jun;2(3):197–218. doi: 10.3109/01677068509100150. [DOI] [PubMed] [Google Scholar]
  5. Engelman A., Rosenberg N. Isolation of temperature-sensitive Abelson virus mutants by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8021–8025. doi: 10.1073/pnas.84.22.8021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gao F. B., Carson C. C., Levine T., Keene J. D. Selection of a subset of mRNAs from combinatorial 3' untranslated region libraries using neuronal RNA-binding protein Hel-N1. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):11207–11211. doi: 10.1073/pnas.91.23.11207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Good P. J. A conserved family of elav-like genes in vertebrates. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4557–4561. doi: 10.1073/pnas.92.10.4557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hoffman D. W., Query C. C., Golden B. L., White S. W., Keene J. D. RNA-binding domain of the A protein component of the U1 small nuclear ribonucleoprotein analyzed by NMR spectroscopy is structurally similar to ribosomal proteins. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2495–2499. doi: 10.1073/pnas.88.6.2495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Homyk T., Jr, Isono K., Pak W. L. Developmental and physiological analysis of a conditional mutation affecting photoreceptor and optic lobe development in Drosophila melanogaster. J Neurogenet. 1985 Nov;2(5):309–324. doi: 10.3109/01677068509102326. [DOI] [PubMed] [Google Scholar]
  10. Homyk T., Jr, Szidonya J., Suzuki D. T. Behavioral mutants of Drosophila melanogaster. III. Isolation and mapping of mutations by direct visual observations of behavioral phenotypes. Mol Gen Genet. 1980;177(4):553–565. doi: 10.1007/BF00272663. [DOI] [PubMed] [Google Scholar]
  11. Johnson D. R., Duronio R. J., Langner C. A., Rudnick D. A., Gordon J. I. Genetic and biochemical studies of a mutant Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase, nmt72pLeu99-->Pro, that produces temperature-sensitive myristic acid auxotrophy. J Biol Chem. 1993 Jan 5;268(1):483–494. [PubMed] [Google Scholar]
  12. Karess R. E., Rubin G. M. Analysis of P transposable element functions in Drosophila. Cell. 1984 Aug;38(1):135–146. doi: 10.1016/0092-8674(84)90534-8. [DOI] [PubMed] [Google Scholar]
  13. Kenan D. J., Query C. C., Keene J. D. RNA recognition: towards identifying determinants of specificity. Trends Biochem Sci. 1991 Jun;16(6):214–220. doi: 10.1016/0968-0004(91)90088-d. [DOI] [PubMed] [Google Scholar]
  14. Kim Y. J., Baker B. S. The Drosophila gene rbp9 encodes a protein that is a member of a conserved group of putative RNA binding proteins that are nervous system-specific in both flies and humans. J Neurosci. 1993 Mar;13(3):1045–1056. doi: 10.1523/JNEUROSCI.13-03-01045.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. King P. H., Levine T. D., Fremeau R. T., Jr, Keene J. D. Mammalian homologs of Drosophila ELAV localized to a neuronal subset can bind in vitro to the 3' UTR of mRNA encoding the Id transcriptional repressor. J Neurosci. 1994 Apr;14(4):1943–1952. doi: 10.1523/JNEUROSCI.14-04-01943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee A. L., Kanaar R., Rio D. C., Wemmer D. E. Resonance assignments and solution structure of the second RNA-binding domain of sex-lethal determined by multidimensional heteronuclear magnetic resonance. Biochemistry. 1994 Nov 22;33(46):13775–13786. doi: 10.1021/bi00250a031. [DOI] [PubMed] [Google Scholar]
  17. Lo P. C., Roy D., Mount S. M. Suppressor U1 snRNAs in Drosophila. Genetics. 1994 Oct;138(2):365–378. doi: 10.1093/genetics/138.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Masai I., Hotta Y. Genomic organization of a Drosophila phospholipase C, norpA, and molecular lesions in two temperature-sensitive mutants. J Biochem. 1991 Jun;109(6):867–871. doi: 10.1093/oxfordjournals.jbchem.a123472. [DOI] [PubMed] [Google Scholar]
  19. Mount S. M., Burks C., Hertz G., Stormo G. D., White O., Fields C. Splicing signals in Drosophila: intron size, information content, and consensus sequences. Nucleic Acids Res. 1992 Aug 25;20(16):4255–4262. doi: 10.1093/nar/20.16.4255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mullins M. C., Rubin G. M. Isolation of temperature-sensitive mutations of the tyrosine kinase receptor sevenless (sev) in Drosophila and their use in determining its time of action. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9387–9391. doi: 10.1073/pnas.88.21.9387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nagai K., Oubridge C., Jessen T. H., Li J., Evans P. R. Crystal structure of the RNA-binding domain of the U1 small nuclear ribonucleoprotein A. Nature. 1990 Dec 6;348(6301):515–520. doi: 10.1038/348515a0. [DOI] [PubMed] [Google Scholar]
  22. Oubridge C., Ito N., Evans P. R., Teo C. H., Nagai K. Crystal structure at 1.92 A resolution of the RNA-binding domain of the U1A spliceosomal protein complexed with an RNA hairpin. Nature. 1994 Dec 1;372(6505):432–438. doi: 10.1038/372432a0. [DOI] [PubMed] [Google Scholar]
  23. Perron M., Théodore L., Wegnez M. Isolation and embryonic expression of Xel-1, a nervous system-specific Xenopus gene related to the elav gene family. Mech Dev. 1995 Jun;51(2-3):235–249. doi: 10.1016/0925-4773(95)00368-1. [DOI] [PubMed] [Google Scholar]
  24. Pirrotta V. Vectors for P-mediated transformation in Drosophila. Biotechnology. 1988;10:437–456. doi: 10.1016/b978-0-409-90042-2.50028-3. [DOI] [PubMed] [Google Scholar]
  25. Rio D. C. Splicing of pre-mRNA: mechanism, regulation and role in development. Curr Opin Genet Dev. 1993 Aug;3(4):574–584. doi: 10.1016/0959-437x(93)90093-5. [DOI] [PubMed] [Google Scholar]
  26. Robinow S., Campos A. R., Yao K. M., White K. The elav gene product of Drosophila, required in neurons, has three RNP consensus motifs. Science. 1988 Dec 16;242(4885):1570–1572. doi: 10.1126/science.3144044. [DOI] [PubMed] [Google Scholar]
  27. Robinow S., White K. Characterization and spatial distribution of the ELAV protein during Drosophila melanogaster development. J Neurobiol. 1991 Jul;22(5):443–461. doi: 10.1002/neu.480220503. [DOI] [PubMed] [Google Scholar]
  28. Sakai K., Gofuku M., Kitagawa Y., Ogasawara T., Hirose G., Yamazaki M., Koh C. S., Yanagisawa N., Steinman L. A hippocampal protein associated with paraneoplastic neurologic syndrome and small cell lung carcinoma. Biochem Biophys Res Commun. 1994 Mar 30;199(3):1200–1208. doi: 10.1006/bbrc.1994.1358. [DOI] [PubMed] [Google Scholar]
  29. Scherly D., Dathan N. A., Boelens W., van Venrooij W. J., Mattaj I. W. The U2B'' RNP motif as a site of protein-protein interaction. EMBO J. 1990 Nov;9(11):3675–3681. doi: 10.1002/j.1460-2075.1990.tb07579.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Th'ng J. P., Wright P. S., Hamaguchi J., Lee M. G., Norbury C. J., Nurse P., Bradbury E. M. The FT210 cell line is a mouse G2 phase mutant with a temperature-sensitive CDC2 gene product. Cell. 1990 Oct 19;63(2):313–324. doi: 10.1016/0092-8674(90)90164-a. [DOI] [PubMed] [Google Scholar]
  31. Thummel C. S., Boulet A. M., Lipshitz H. D. Vectors for Drosophila P-element-mediated transformation and tissue culture transfection. Gene. 1988 Dec 30;74(2):445–456. doi: 10.1016/0378-1119(88)90177-1. [DOI] [PubMed] [Google Scholar]
  32. Valle R. P., Morch M. D. Stop making sense: or Regulation at the level of termination in eukaryotic protein synthesis. FEBS Lett. 1988 Aug 1;235(1-2):1–15. doi: 10.1016/0014-5793(88)81225-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Xue F., Cooley L. kelch encodes a component of intercellular bridges in Drosophila egg chambers. Cell. 1993 Mar 12;72(5):681–693. doi: 10.1016/0092-8674(93)90397-9. [DOI] [PubMed] [Google Scholar]
  34. Yao K. M., Samson M. L., Reeves R., White K. Gene elav of Drosophila melanogaster: a prototype for neuronal-specific RNA binding protein gene family that is conserved in flies and humans. J Neurobiol. 1993 Jun;24(6):723–739. doi: 10.1002/neu.480240604. [DOI] [PubMed] [Google Scholar]
  35. Yao K. M., White K. Organizational analysis of elav gene and functional analysis of ELAV protein of Drosophila melanogaster and Drosophila virilis. Mol Cell Biol. 1991 Jun;11(6):2994–3000. doi: 10.1128/mcb.11.6.2994. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES