Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Aug;14(8):5412–5424. doi: 10.1128/mcb.14.8.5412

La proteins from Drosophila melanogaster and Saccharomyces cerevisiae: a yeast homolog of the La autoantigen is dispensable for growth.

C J Yoo 1, S L Wolin 1
PMCID: PMC359060  PMID: 8035818

Abstract

The human autoantigen La is a 50-kDa protein which binds to the 3' termini of virtually all nascent polymerase III transcripts. Experiments with mammalian transcription extracts have led to the proposal that the La protein is required for multiple rounds of transcription by RNA polymerase III (E. Gottlieb and J. A. Steitz, EMBO J. 8:851-861, 1989; R. J. Maraia, D. J. Kenan, and J. D. Keene, Mol. Cell. Biol. 14:2147-2158, 1994). Although La protein homologs have been identified in a variety of vertebrate species, the protein has not been identified in invertebrates. In order to begin a genetic analysis of La protein function, we have characterized homologs of the La protein in the fruit fly Drosophila melanogaster and the yeast Saccharomyces cerevisiae. We show that both the Drosophila and yeast La proteins are bound to precursors of polymerase III RNAs in vivo. The Drosophila and yeast proteins resemble the human La protein in their biochemical properties, as both proteins can be partially purified from cells by a procedure previously devised to purify the human protein. Similarly to vertebrate La proteins, the Drosophila and yeast homologs preferentially bind RNAs that terminate with a 3' hydroxyl. Despite the fact that the La protein is conserved between humans and Saccharomyces cerevisiae, yeast cells containing a null allele of the gene encoding the La protein are viable, suggesting that another protein(s) plays a functionally redundant role.

Full text

PDF
5412

Images in this article

5417Image
on p.5417
5418Image
on p.5418
5419Image
on p.5419
5420Image
on p.5420
5421Image
on p.5421
5421Image
on p.5421
5422Image
on p.5422

Selected References

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

  1. Adams A. E., Cooper J. A., Drubin D. G. Unexpected combinations of null mutations in genes encoding the actin cytoskeleton are lethal in yeast. Mol Biol Cell. 1993 May;4(5):459–468. doi: 10.1091/mbc.4.5.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Bender A., Pringle J. R. Use of a screen for synthetic lethal and multicopy suppressee mutants to identify two new genes involved in morphogenesis in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Mar;11(3):1295–1305. doi: 10.1128/mcb.11.3.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Black D. L., Pinto A. L. U5 small nuclear ribonucleoprotein: RNA structure analysis and ATP-dependent interaction with U4/U6. Mol Cell Biol. 1989 Aug;9(8):3350–3359. doi: 10.1128/mcb.9.8.3350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brow D. A. In vitro transcripts of a yeast variant 5 S rRNA gene exhibit alterations in 3'-end processing and protein binding. J Biol Chem. 1987 Oct 15;262(29):13959–13965. [PubMed] [Google Scholar]
  6. Campbell F. E., Jr, Setzer D. R. Transcription termination by RNA polymerase III: uncoupling of polymerase release from termination signal recognition. Mol Cell Biol. 1992 May;12(5):2260–2272. doi: 10.1128/mcb.12.5.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chambers J. C., Kenan D., Martin B. J., Keene J. D. Genomic structure and amino acid sequence domains of the human La autoantigen. J Biol Chem. 1988 Dec 5;263(34):18043–18051. [PubMed] [Google Scholar]
  8. Chambers J. C., Kurilla M. G., Keene J. D. Association between the 7 S RNA and the lupus La protein varies among cell types. J Biol Chem. 1983 Oct 10;258(19):11438–11441. [PubMed] [Google Scholar]
  9. Chan E. K., Sullivan K. F., Tan E. M. Ribonucleoprotein SS-B/La belongs to a protein family with consensus sequences for RNA-binding. Nucleic Acids Res. 1989 Mar 25;17(6):2233–2244. doi: 10.1093/nar/17.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cozzarelli N. R., Gerrard S. P., Schlissel M., Brown D. D., Bogenhagen D. F. Purified RNA polymerase III accurately and efficiently terminates transcription of 5S RNA genes. Cell. 1983 Oct;34(3):829–835. doi: 10.1016/0092-8674(83)90540-8. [DOI] [PubMed] [Google Scholar]
  11. Driever W., Nüsslein-Volhard C. A gradient of bicoid protein in Drosophila embryos. Cell. 1988 Jul 1;54(1):83–93. doi: 10.1016/0092-8674(88)90182-1. [DOI] [PubMed] [Google Scholar]
  12. England T. E., Bruce A. G., Uhlenbeck O. C. Specific labeling of 3' termini of RNA with T4 RNA ligase. Methods Enzymol. 1980;65(1):65–74. doi: 10.1016/s0076-6879(80)65011-3. [DOI] [PubMed] [Google Scholar]
  13. Felici F., Cesareni G., Hughes J. M. The most abundant small cytoplasmic RNA of Saccharomyces cerevisiae has an important function required for normal cell growth. Mol Cell Biol. 1989 Aug;9(8):3260–3268. doi: 10.1128/mcb.9.8.3260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Francoeur A. M., Gritzmacher C. A., Peebles C. L., Reese R. T., Tan E. M. Synthesis of small nuclear ribonucleoprotein particles by the malarial parasite Plasmodium falciparum. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3635–3639. doi: 10.1073/pnas.82.11.3635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Hann B. C., Walter P. The signal recognition particle in S. cerevisiae. Cell. 1991 Oct 4;67(1):131–144. doi: 10.1016/0092-8674(91)90577-l. [DOI] [PubMed] [Google Scholar]
  18. Hendrick J. P., Wolin S. L., Rinke J., Lerner M. R., Steitz J. A. Ro small cytoplasmic ribonucleoproteins are a subclass of La ribonucleoproteins: further characterization of the Ro and La small ribonucleoproteins from uninfected mammalian cells. Mol Cell Biol. 1981 Dec;1(12):1138–1149. doi: 10.1128/mcb.1.12.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jacq B., Jourdan R., Jordan B. R. Structure and processing of precursor 5 S RNA in Drosophila melanogaster. J Mol Biol. 1977 Dec 15;117(3):785–795. doi: 10.1016/0022-2836(77)90069-9. [DOI] [PubMed] [Google Scholar]
  20. Karpen G. H., Spradling A. C. Analysis of subtelomeric heterochromatin in the Drosophila minichromosome Dp1187 by single P element insertional mutagenesis. Genetics. 1992 Nov;132(3):737–753. doi: 10.1093/genetics/132.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Koonin E. V., Bork P., Sander C. Yeast chromosome III: new gene functions. EMBO J. 1994 Feb 1;13(3):493–503. doi: 10.1002/j.1460-2075.1994.tb06287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lerner M. R., Andrews N. C., Miller G., Steitz J. A. Two small RNAs encoded by Epstein-Barr virus and complexed with protein are precipitated by antibodies from patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1981 Feb;78(2):805–809. doi: 10.1073/pnas.78.2.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lerner M. R., Boyle J. A., Hardin J. A., Steitz J. A. Two novel classes of small ribonucleoproteins detected by antibodies associated with lupus erythematosus. Science. 1981 Jan 23;211(4480):400–402. doi: 10.1126/science.6164096. [DOI] [PubMed] [Google Scholar]
  25. Maraia R. J., Kenan D. J., Keene J. D. Eukaryotic transcription termination factor La mediates transcript release and facilitates reinitiation by RNA polymerase III. Mol Cell Biol. 1994 Mar;14(3):2147–2158. doi: 10.1128/mcb.14.3.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mathews M. B., Francoeur A. M. La antigen recognizes and binds to the 3'-oligouridylate tail of a small RNA. Mol Cell Biol. 1984 Jun;4(6):1134–1140. doi: 10.1128/mcb.4.6.1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Meerovitch K., Svitkin Y. V., Lee H. S., Lejbkowicz F., Kenan D. J., Chan E. K., Agol V. I., Keene J. D., Sonenberg N. La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate. J Virol. 1993 Jul;67(7):3798–3807. doi: 10.1128/jvi.67.7.3798-3807.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Myer V. E., Lee S. I., Steitz J. A. Viral small nuclear ribonucleoproteins bind a protein implicated in messenger RNA destabilization. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1296–1300. doi: 10.1073/pnas.89.4.1296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nagiec M. M., Wells G. B., Lester R. L., Dickson R. C. A suppressor gene that enables Saccharomyces cerevisiae to grow without making sphingolipids encodes a protein that resembles an Escherichia coli fatty acyltransferase. J Biol Chem. 1993 Oct 15;268(29):22156–22163. [PubMed] [Google Scholar]
  30. Oliver S. G., van der Aart Q. J., Agostoni-Carbone M. L., Aigle M., Alberghina L., Alexandraki D., Antoine G., Anwar R., Ballesta J. P., Benit P. The complete DNA sequence of yeast chromosome III. Nature. 1992 May 7;357(6373):38–46. doi: 10.1038/357038a0. [DOI] [PubMed] [Google Scholar]
  31. Piper P. W., Bellatin J. A., Lockheart A. Altered maturation of sequences at the 3' terminus of 5S gene transcripts in a Saccharomyces cerevisiae mutant that lacks a RNA processing endonuclease. EMBO J. 1983;2(3):353–359. doi: 10.1002/j.1460-2075.1983.tb01430.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Piper P. W., Patel N., Lockheart A. Processing of the 3' sequence extensions upon the 5S rRNA of a mutant yeast in Xenopus laevis germinal vesicle extract. Eur J Biochem. 1984 May 15;141(1):115–118. doi: 10.1111/j.1432-1033.1984.tb08165.x. [DOI] [PubMed] [Google Scholar]
  33. Preiser P., Vasisht V., Birk A., Levinger L. Poly(U)-binding protein inhibits Drosophila pre-5 S RNA 3'-exonuclease digestion. J Biol Chem. 1993 Jun 5;268(16):11553–11557. [PubMed] [Google Scholar]
  34. 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]
  35. Query C. C., Bentley R. C., Keene J. D. A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70K U1 snRNP protein. Cell. 1989 Apr 7;57(1):89–101. doi: 10.1016/0092-8674(89)90175-x. [DOI] [PubMed] [Google Scholar]
  36. Reddy R., Henning D., Tan E., Busch H. Identification of a La protein binding site in a RNA polymerase III transcript (4.5 I RNA). J Biol Chem. 1983 Jul 10;258(13):8352–8356. [PubMed] [Google Scholar]
  37. Rinke J., Steitz J. A. Association of the lupus antigen La with a subset of U6 snRNA molecules. Nucleic Acids Res. 1985 Apr 11;13(7):2617–2629. doi: 10.1093/nar/13.7.2617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rinke J., Steitz J. A. Precursor molecules of both human 5S ribosomal RNA and transfer RNAs are bound by a cellular protein reactive with anti-La lupus antibodies. Cell. 1982 May;29(1):149–159. doi: 10.1016/0092-8674(82)90099-x. [DOI] [PubMed] [Google Scholar]
  39. Rosa M. D., Gottlieb E., Lerner M. R., Steitz J. A. Striking similarities are exhibited by two small Epstein-Barr virus-encoded ribonucleic acids and the adenovirus-associated ribonucleic acids VAI and VAII. Mol Cell Biol. 1981 Sep;1(9):785–796. doi: 10.1128/mcb.1.9.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scherly D., Stutz F., Lin-Marq N., Clarkson S. G. La proteins from Xenopus laevis. cDNA cloning and developmental expression. J Mol Biol. 1993 May 20;231(2):196–204. doi: 10.1006/jmbi.1993.1275. [DOI] [PubMed] [Google Scholar]
  41. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Terns M. P., Lund E., Dahlberg J. E. 3'-end-dependent formation of U6 small nuclear ribonucleoprotein particles in Xenopus laevis oocyte nuclei. Mol Cell Biol. 1992 Jul;12(7):3032–3040. doi: 10.1128/mcb.12.7.3032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Topfer F., Gordon T., McCluskey J. Characterization of the mouse autoantigen La (SS-B). Identification of conserved RNA-binding motifs, a putative ATP binding site and reactivity of recombinant protein with poly(U) and human autoantibodies. J Immunol. 1993 Apr 1;150(7):3091–3100. [PubMed] [Google Scholar]
  45. Watson J. B., Chandler D. W., Gralla J. D. Specific termination of in vitro transcription by calf thymus RNA polymerase III. Nucleic Acids Res. 1984 Jul 11;12(13):5369–5384. doi: 10.1093/nar/12.13.5369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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]
  47. Yisraeli J. K., Melton D. A. Synthesis of long, capped transcripts in vitro by SP6 and T7 RNA polymerases. Methods Enzymol. 1989;180:42–50. doi: 10.1016/0076-6879(89)80090-4. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES