Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1999 Jan;151(1):143–150. doi: 10.1093/genetics/151.1.143

The levels of the RoRNP-associated Y RNA are dependent upon the presence of ROP-1, the Caenorhabditis elegans Ro60 protein.

J C Labbé 1, S Hekimi 1, L A Rokeach 1
PMCID: PMC1460465  PMID: 9872955

Abstract

The Ro ribonucleoproteins (RoRNP) consist of at least one major protein of 60 kD, Ro60, and one small associated RNA, designated Y RNA. Although RoRNP have been found in all vertebrate species examined so far, their function remains unknown. The Caenorhabditis elegans rop-1 gene previously has been identified as encoding a Ro60 homologue. We report here the phenotypic characterization of a C. elegans strain in which rop-1 has been disrupted. This is the first report regarding the inactivation of a major RoRNP constituent in any organism. The rop-1 mutant worms display no visible defects. However, at the molecular level, the disruption of rop-1 results in a dramatic decrease in the levels of the ROP-1-associated RNA (CeY RNA). Moreover, transgenic expression of wild-type rop-1 partially rescues the levels of CeY RNA. Considering that transgenes are poorly expressed in the germline, the fact that the rescue is only partial is most likely related to the high abundance of the CeY RNA in the adult germline and in embryos. The developmental expression pattern and localization of CeY RNA suggest a role for this molecule during embryogenesis. We conclude that, under laboratory culture conditions, ROP-1 does not play a crucial role in C. elegans.

Full Text

The Full Text of this article is available as a PDF (188.1 KB).

Selected References

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

  1. Ali N., Siddiqui A. The La antigen binds 5' noncoding region of the hepatitis C virus RNA in the context of the initiator AUG codon and stimulates internal ribosome entry site-mediated translation. Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2249–2254. doi: 10.1073/pnas.94.6.2249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Austin J., Kimble J. glp-1 is required in the germ line for regulation of the decision between mitosis and meiosis in C. elegans. Cell. 1987 Nov 20;51(4):589–599. doi: 10.1016/0092-8674(87)90128-0. [DOI] [PubMed] [Google Scholar]
  3. Bai C., Li Z., Tolias P. P. Developmental characterization of a Drosophila RNA-binding protein homologous to the human systemic lupus erythematosus-associated La/SS-B autoantigen. Mol Cell Biol. 1994 Aug;14(8):5123–5129. doi: 10.1128/mcb.14.8.5123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barton M. K., Schedl T. B., Kimble J. Gain-of-function mutations of fem-3, a sex-determination gene in Caenorhabditis elegans. Genetics. 1987 Jan;115(1):107–119. doi: 10.1093/genetics/115.1.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Belsham G. J., Sonenberg N., Svitkin Y. V. The role of the La autoantigen in internal initiation. Curr Top Microbiol Immunol. 1995;203:85–98. doi: 10.1007/978-3-642-79663-0_4. [DOI] [PubMed] [Google Scholar]
  6. Boire G., Craft J. Biochemical and immunological heterogeneity of the Ro ribonucleoprotein particles. Analysis with sera specific for the RohY5 particle. J Clin Invest. 1989 Jul;84(1):270–279. doi: 10.1172/JCI114150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Boire G., Gendron M., Monast N., Bastin B., Ménard H. A. Purification of antigenically intact Ro ribonucleoproteins; biochemical and immunological evidence that the 52-kD protein is not a Ro protein. Clin Exp Immunol. 1995 Jun;100(3):489–498. doi: 10.1111/j.1365-2249.1995.tb03728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Chan E. K., Hamel J. C., Buyon J. P., Tan E. M. Molecular definition and sequence motifs of the 52-kD component of human SS-A/Ro autoantigen. J Clin Invest. 1991 Jan;87(1):68–76. doi: 10.1172/JCI115003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Deutscher S. L., Harley J. B., Keene J. D. Molecular analysis of the 60-kDa human Ro ribonucleoprotein. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9479–9483. doi: 10.1073/pnas.85.24.9479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ewbank J. J., Barnes T. M., Lakowski B., Lussier M., Bussey H., Hekimi S. Structural and functional conservation of the Caenorhabditis elegans timing gene clk-1. Science. 1997 Feb 14;275(5302):980–983. doi: 10.1126/science.275.5302.980. [DOI] [PubMed] [Google Scholar]
  12. Fan H., Goodier J. L., Chamberlain J. R., Engelke D. R., Maraia R. J. 5' processing of tRNA precursors can Be modulated by the human La antigen phosphoprotein. Mol Cell Biol. 1998 Jun;18(6):3201–3211. doi: 10.1128/mcb.18.6.3201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fan H., Sakulich A. L., Goodier J. L., Zhang X., Qin J., Maraia R. J. Phosphorylation of the human La antigen on serine 366 can regulate recycling of RNA polymerase III transcription complexes. Cell. 1997 Mar 7;88(5):707–715. doi: 10.1016/s0092-8674(00)81913-3. [DOI] [PubMed] [Google Scholar]
  14. Farris A. D., O'Brien C. A., Harley J. B. Y3 is the most conserved small RNA component of Ro ribonucleoprotein complexes in vertebrate species. Gene. 1995 Mar 10;154(2):193–198. doi: 10.1016/0378-1119(94)00823-b. [DOI] [PubMed] [Google Scholar]
  15. Farris A. D., Puvion-Dutilleul F., Puvion E., Harley J. B., Lee L. A. The ultrastructural localization of 60-kDa Ro protein and human cytoplasmic RNAs: association with novel electron-dense bodies. Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3040–3045. doi: 10.1073/pnas.94.7.3040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ferguson K. C., Heid P. J., Rothman J. H. The SL1 trans-spliced leader RNA performs an essential embryonic function in Caenorhabditis elegans that can also be supplied by SL2 RNA. Genes Dev. 1996 Jun 15;10(12):1543–1556. doi: 10.1101/gad.10.12.1543. [DOI] [PubMed] [Google Scholar]
  17. Fire A., Harrison S. W., Dixon D. A modular set of lacZ fusion vectors for studying gene expression in Caenorhabditis elegans. Gene. 1990 Sep 14;93(2):189–198. doi: 10.1016/0378-1119(90)90224-f. [DOI] [PubMed] [Google Scholar]
  18. Fire A. Histochemical techniques for locating Escherichia coli beta-galactosidase activity in transgenic organisms. Genet Anal Tech Appl. 1992 Oct-Dec;9(5-6):151–158. doi: 10.1016/1050-3862(92)90042-4. [DOI] [PubMed] [Google Scholar]
  19. Gottlieb E., Steitz J. A. Function of the mammalian La protein: evidence for its action in transcription termination by RNA polymerase III. EMBO J. 1989 Mar;8(3):851–861. doi: 10.1002/j.1460-2075.1989.tb03446.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gottlieb E., Steitz J. A. The RNA binding protein La influences both the accuracy and the efficiency of RNA polymerase III transcription in vitro. EMBO J. 1989 Mar;8(3):841–850. doi: 10.1002/j.1460-2075.1989.tb03445.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Green C. D., Long K. S., Shi H., Wolin S. L. Binding of the 60-kDa Ro autoantigen to Y RNAs: evidence for recognition in the major groove of a conserved helix. RNA. 1998 Jul;4(7):750–765. doi: 10.1017/s1355838298971667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hühn P., Pruijn G. J., van Venrooij W. J., Bachmann M. Characterization of the autoantigen La (SS-B) as a dsRNA unwinding enzyme. Nucleic Acids Res. 1997 Jan 15;25(2):410–416. doi: 10.1093/nar/25.2.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Itoh K., Itoh Y., Frank M. B. Protein heterogeneity in the human Ro/SSA ribonucleoproteins. The 52- and 60-kD Ro/SSA autoantigens are encoded by separate genes. J Clin Invest. 1991 Jan;87(1):177–186. doi: 10.1172/JCI114968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kelekar A., Saitta M. R., Keene J. D. Molecular composition of Ro small ribonucleoprotein complexes in human cells. Intracellular localization of the 60- and 52-kD proteins. J Clin Invest. 1994 Apr;93(4):1637–1644. doi: 10.1172/JCI117145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Krause M., Wild M., Rosenzweig B., Hirsh D. Wild-type and mutant actin genes in Caenorhabditis elegans. J Mol Biol. 1989 Aug 5;208(3):381–392. doi: 10.1016/0022-2836(89)90503-2. [DOI] [PubMed] [Google Scholar]
  26. Labbé J. C., Jannatipour M., Rokeach L. A. The Caenorhabditis elegans rop-1 gene encodes the homologue of the human 60-kDa Ro autoantigen. Gene. 1995 Dec 29;167(1-2):227–231. doi: 10.1016/0378-1119(95)00695-8. [DOI] [PubMed] [Google Scholar]
  27. Maraia R. J. Transcription termination factor La is also an initiation factor for RNA polymerase III. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3383–3387. doi: 10.1073/pnas.93.8.3383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mello C., Fire A. DNA transformation. Methods Cell Biol. 1995;48:451–482. [PubMed] [Google Scholar]
  29. O'Brien C. A., Margelot K., Wolin S. L. Xenopus Ro ribonucleoproteins: members of an evolutionarily conserved class of cytoplasmic ribonucleoproteins. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7250–7254. doi: 10.1073/pnas.90.15.7250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Brien C. A., Wolin S. L. A possible role for the 60-kD Ro autoantigen in a discard pathway for defective 5S rRNA precursors. Genes Dev. 1994 Dec 1;8(23):2891–2903. doi: 10.1101/gad.8.23.2891. [DOI] [PubMed] [Google Scholar]
  31. Pruijn G. J., Slobbe R. L., van Venrooij W. J. Analysis of protein--RNA interactions within Ro ribonucleoprotein complexes. Nucleic Acids Res. 1991 Oct 11;19(19):5173–5180. doi: 10.1093/nar/19.19.5173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pruijn G. J., Wingens P. A., Peters S. L., Thijssen J. P., van Venrooij W. J. Ro RNP associated Y RNAs are highly conserved among mammals. Biochim Biophys Acta. 1993 Dec 14;1216(3):395–401. doi: 10.1016/0167-4781(93)90006-y. [DOI] [PubMed] [Google Scholar]
  33. Rokeach L. A., Haselby J. A., Hoch S. O. High-level bacterial expression, purification and characterization of human calreticulin. Protein Eng. 1991 Dec;4(8):981–987. doi: 10.1093/protein/4.8.981. [DOI] [PubMed] [Google Scholar]
  34. Shi H., O'Brien C. A., Van Horn D. J., Wolin S. L. A misfolded form of 5S rRNA is complexed with the Ro and La autoantigens. RNA. 1996 Aug;2(8):769–784. [PMC free article] [PubMed] [Google Scholar]
  35. Slobbe R. L., Pluk W., van Venrooij W. J., Pruijn G. J. Ro ribonucleoprotein assembly in vitro. Identification of RNA-protein and protein-protein interactions. J Mol Biol. 1992 Sep 20;227(2):361–366. doi: 10.1016/0022-2836(92)90890-v. [DOI] [PubMed] [Google Scholar]
  36. Stefano J. E. Purified lupus antigen La recognizes an oligouridylate stretch common to the 3' termini of RNA polymerase III transcripts. Cell. 1984 Jan;36(1):145–154. doi: 10.1016/0092-8674(84)90083-7. [DOI] [PubMed] [Google Scholar]
  37. Tan E. M. Molecular biology of nuclear autoantigens. Adv Nephrol Necker Hosp. 1993;22:213–236. [PubMed] [Google Scholar]
  38. Tseng C. E., Chan E. K., Miranda E., Gross M., Di Donato F., Buyon J. P. The 52-kd protein as a target of intermolecular spreading of the immune response to components of the SS-A/Ro-SS-B/La complex. Arthritis Rheum. 1997 May;40(5):936–944. doi: 10.1002/art.1780400523. [DOI] [PubMed] [Google Scholar]
  39. Van Horn D. J., Eisenberg D., O'Brien C. A., Wolin S. L. Caenorhabditis elegans embryos contain only one major species of Ro RNP. RNA. 1995 May;1(3):293–303. [PMC free article] [PubMed] [Google Scholar]
  40. Van Horn D. J., Yoo C. J., Xue D., Shi H., Wolin S. L. The La protein in Schizosaccharomyces pombe: a conserved yet dispensable phosphoprotein that functions in tRNA maturation. RNA. 1997 Dec;3(12):1434–1443. [PMC free article] [PubMed] [Google Scholar]
  41. Wang D., Buyon J. P., Chan E. K. Cloning and expression of mouse 60 kDa ribonucleoprotein SS-A/Ro. Mol Biol Rep. 1996;23(3-4):205–210. doi: 10.1007/BF00351170. [DOI] [PubMed] [Google Scholar]
  42. Wolffe A. P., Brown D. D. Developmental regulation of two 5S ribosomal RNA genes. Science. 1988 Sep 23;241(4873):1626–1632. doi: 10.1126/science.241.4873.1626. [DOI] [PubMed] [Google Scholar]
  43. Wolin S. L., Steitz J. A. Genes for two small cytoplasmic Ro RNAs are adjacent and appear to be single-copy in the human genome. Cell. 1983 Mar;32(3):735–744. doi: 10.1016/0092-8674(83)90059-4. [DOI] [PubMed] [Google Scholar]
  44. Wolin S. L., Steitz J. A. The Ro small cytoplasmic ribonucleoproteins: identification of the antigenic protein and its binding site on the Ro RNAs. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1996–2000. doi: 10.1073/pnas.81.7.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yoo C. J., Wolin S. L. The yeast La protein is required for the 3' endonucleolytic cleavage that matures tRNA precursors. Cell. 1997 May 2;89(3):393–402. doi: 10.1016/s0092-8674(00)80220-2. [DOI] [PubMed] [Google Scholar]
  46. Youinou P., Adler Y., Muller S., Lamour A., Baron D., Humbel R. L. Anti-Ro(SSA) and anti-La(SSB) antibodies in autoimmune rheumatic diseases. Clin Rev Allergy. 1994 Fall;12(3):253–274. doi: 10.1007/BF02802321. [DOI] [PubMed] [Google Scholar]
  47. Zwaal R. R., Broeks A., van Meurs J., Groenen J. T., Plasterk R. H. Target-selected gene inactivation in Caenorhabditis elegans by using a frozen transposon insertion mutant bank. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7431–7435. doi: 10.1073/pnas.90.16.7431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. van Gelder C. W., Thijssen J. P., Klaassen E. C., Sturchler C., Krol A., van Venrooij W. J., Pruijn G. J. Common structural features of the Ro RNP associated hY1 and hY5 RNAs. Nucleic Acids Res. 1994 Jul 11;22(13):2498–2506. doi: 10.1093/nar/22.13.2498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. van Venrooij W. J., Slobbe R. L., Pruijn G. J. Structure and function of La and Ro RNPs. Mol Biol Rep. 1993 Aug;18(2):113–119. doi: 10.1007/BF00986765. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES