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. 1997 Oct;17(10):5823–5832. doi: 10.1128/mcb.17.10.5823

A carboxy-terminal basic region controls RNA polymerase III transcription factor activity of human La protein.

J L Goodier 1, H Fan 1, R J Maraia 1
PMCID: PMC232430  PMID: 9315640

Abstract

Human La protein has been shown to serve as a transcription factor for RNA polymerase III (pol III) by facilitating transcription termination and recycling of transcription complexes. In addition, La binds to the 3' oligo(U) ends common to all nascent pol III transcripts, and in the case of B1-Alu RNA, protects it from 3'-end processing (R. J. Maraia, D. J. Kenan, and J. D. Keene, Mol. Cell. Biol. 14:2147-2158, 1994). Others have previously dissected the La protein into an N-terminal domain that binds RNA and a C-terminal domain that does not. Here, deletion and substitution mutants of La were examined for general RNA binding, RNA 3'-end protection, and transcription factor activity. Although some La mutants altered in a C-terminal basic region bind RNA in mobility shift assays, they are defective in RNA 3'-end protection and do not support transcription, while one C-terminal substitution mutant is defective only in transcription. Moreover, a C-terminal fragment lacking RNA binding activity appears able to support low levels of transcription by pol III. While efficient multiround transcription is supported only by mutants that bind RNA and contain a C-terminal basic region. These analyses indicate that RNA binding contributes to but is not sufficient for La transcription factor activity and that the C-terminal domain plays a role in transcription that is distinguishable from simple RNA binding. The transcription factor activity of La can be reversibly inhibited by RNA, suggesting the potential for feedback inhibition of pol III transcription.

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Selected References

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  1. Adeniyi-Jones S., Zasloff M. Transcription, processing and nuclear transport of a B1 Alu RNA species complementary to an intron of the murine alpha-fetoprotein gene. Nature. 1985 Sep 5;317(6032):81–84. doi: 10.1038/317081a0. [DOI] [PubMed] [Google Scholar]
  2. Allison D. S., Hall B. D. Effects of alterations in the 3' flanking sequence on in vivo and in vitro expression of the yeast SUP4-o tRNATyr gene. EMBO J. 1985 Oct;4(10):2657–2664. doi: 10.1002/j.1460-2075.1985.tb03984.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birney E., Kumar S., Krainer A. R. Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors. Nucleic Acids Res. 1993 Dec 25;21(25):5803–5816. doi: 10.1093/nar/21.25.5803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bogenhagen D. F., Brown D. D. Nucleotide sequences in Xenopus 5S DNA required for transcription termination. Cell. 1981 Apr;24(1):261–270. doi: 10.1016/0092-8674(81)90522-5. [DOI] [PubMed] [Google Scholar]
  5. Bogenhagen D. F., Wormington W. M., Brown D. D. Stable transcription complexes of Xenopus 5S RNA genes: a means to maintain the differentiated state. Cell. 1982 Feb;28(2):413–421. doi: 10.1016/0092-8674(82)90359-2. [DOI] [PubMed] [Google Scholar]
  6. Brow D. A., Geiduschek E. P. Modulation of yeast 5 S rRNA synthesis in vitro by ribosomal protein YL3. A possible regulatory loop. J Biol Chem. 1987 Oct 15;262(29):13953–13958. [PubMed] [Google Scholar]
  7. Chan E. K., Tan E. M. The small nuclear ribonucleoprotein SS-B/La binds RNA with a conserved protease-resistant domain of 28 kilodaltons. Mol Cell Biol. 1987 Jul;7(7):2588–2591. doi: 10.1128/mcb.7.7.2588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang D. Y., Maraia R. J. A cellular protein binds B1 and Alu small cytoplasmic RNAs in vitro. J Biol Chem. 1993 Mar 25;268(9):6423–6428. [PubMed] [Google Scholar]
  9. Chang Y. N., Kenan D. J., Keene J. D., Gatignol A., Jeang K. T. Direct interactions between autoantigen La and human immunodeficiency virus leader RNA. J Virol. 1994 Nov;68(11):7008–7020. doi: 10.1128/jvi.68.11.7008-7020.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Craig A. W., Svitkin Y. V., Lee H. S., Belsham G. J., Sonenberg N. The La autoantigen contains a dimerization domain that is essential for enhancing translation. Mol Cell Biol. 1997 Jan;17(1):163–169. doi: 10.1128/mcb.17.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dean N., Berk A. J. Ordering promoter binding of class III transcription factors TFIIIC1 and TFIIIC2. Mol Cell Biol. 1988 Aug;8(8):3017–3025. doi: 10.1128/mcb.8.8.3017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dean N., Berk A. J. Separation of TFIIIC into two functional components by sequence specific DNA affinity chromatography. Nucleic Acids Res. 1987 Dec 10;15(23):9895–9907. doi: 10.1093/nar/15.23.9895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dieci G., Duimio L., Coda-Zabetta F., Sprague K. U., Ottonello S. A novel RNA polymerase III transcription factor fraction that is not required for template commitment. J Biol Chem. 1993 May 25;268(15):11199–11207. [PubMed] [Google Scholar]
  14. Dieci G., Sentenac A. Facilitated recycling pathway for RNA polymerase III. Cell. 1996 Jan 26;84(2):245–252. doi: 10.1016/s0092-8674(00)80979-4. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  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. Kassavetis G. A., Braun B. R., Nguyen L. H., Geiduschek E. P. S. cerevisiae TFIIIB is the transcription initiation factor proper of RNA polymerase III, while TFIIIA and TFIIIC are assembly factors. Cell. 1990 Jan 26;60(2):235–245. doi: 10.1016/0092-8674(90)90739-2. [DOI] [PubMed] [Google Scholar]
  20. Kassavetis G. A., Nguyen S. T., Kobayashi R., Kumar A., Geiduschek E. P., Pisano M. Cloning, expression, and function of TFC5, the gene encoding the B" component of the Saccharomyces cerevisiae RNA polymerase III transcription factor TFIIIB. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9786–9790. doi: 10.1073/pnas.92.21.9786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kovelman R., Roeder R. G. Sarkosyl defines three intermediate steps in transcription initiation by RNA polymerase III: application to stimulation of transcription by E1A. Genes Dev. 1990 Apr;4(4):646–658. doi: 10.1101/gad.4.4.646. [DOI] [PubMed] [Google Scholar]
  22. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  23. Lagna G., Kovelman R., Sukegawa J., Roeder R. G. Cloning and characterization of an evolutionarily divergent DNA-binding subunit of mammalian TFIIIC. Mol Cell Biol. 1994 May;14(5):3053–3064. doi: 10.1128/mcb.14.5.3053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lassar A. B., Martin P. L., Roeder R. G. Transcription of class III genes: formation of preinitiation complexes. Science. 1983 Nov 18;222(4625):740–748. doi: 10.1126/science.6356356. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Maraia R. J., Chang D. Y., Wolffe A. P., Vorce R. L., Hsu K. The RNA polymerase III terminator used by a B1-Alu element can modulate 3' processing of the intermediate RNA product. Mol Cell Biol. 1992 Apr;12(4):1500–1506. doi: 10.1128/mcb.12.4.1500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Maraia R. J., Sasaki-Tozawa N., Driscoll C. T., Green E. D., Darlington G. J. The human Y4 small cytoplasmic RNA gene is controlled by upstream elements and resides on chromosome 7 with all other hY scRNA genes. Nucleic Acids Res. 1994 Aug 11;22(15):3045–3052. doi: 10.1093/nar/22.15.3045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maraia R. J. The subset of mouse B1 (Alu-equivalent) sequences expressed as small processed cytoplasmic transcripts. Nucleic Acids Res. 1991 Oct 25;19(20):5695–5702. doi: 10.1093/nar/19.20.5695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Maraia R., Zasloff M., Plotz P., Adeniyi-Jones S. Pathway of B1-Alu expression in microinjected oocytes: Xenopus laevis proteins associated with nuclear precursor and processed cytoplasmic RNAs. Mol Cell Biol. 1988 Oct;8(10):4433–4440. doi: 10.1128/mcb.8.10.4433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Rüth J., Conesa C., Dieci G., Lefebvre O., Düsterhöft A., Ottonello S., Sentenac A. A suppressor of mutations in the class III transcription system encodes a component of yeast TFIIIB. EMBO J. 1996 Apr 15;15(8):1941–1949. [PMC free article] [PubMed] [Google Scholar]
  35. Sinn E., Wang Z., Kovelman R., Roeder R. G. Cloning and characterization of a TFIIIC2 subunit (TFIIIC beta) whose presence correlates with activation of RNA polymerase III-mediated transcription by adenovirus E1A expression and serum factors. Genes Dev. 1995 Mar 15;9(6):675–685. doi: 10.1101/gad.9.6.675. [DOI] [PubMed] [Google Scholar]
  36. Sklar V. E., Roeder R. G. Purification and subunit structure of deoxyribonucleic acid-dependent ribonucleic acid polymerase III from the mouse plasmacytoma, MOPC 315. J Biol Chem. 1976 Feb 25;251(4):1064–1073. [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Wang Z., Roeder R. G. Structure and function of a human transcription factor TFIIIB subunit that is evolutionarily conserved and contains both TFIIB- and high-mobility-group protein 2-related domains. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7026–7030. doi: 10.1073/pnas.92.15.7026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang Z., Roeder R. G. TFIIIC1 acts through a downstream region to stabilize TFIIIC2 binding to RNA polymerase III promoters. Mol Cell Biol. 1996 Dec;16(12):6841–6850. doi: 10.1128/mcb.16.12.6841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Willis I. M. RNA polymerase III. Genes, factors and transcriptional specificity. Eur J Biochem. 1993 Feb 15;212(1):1–11. doi: 10.1111/j.1432-1033.1993.tb17626.x. [DOI] [PubMed] [Google Scholar]
  42. Yoo C. J., Wolin S. L. La proteins from Drosophila melanogaster and Saccharomyces cerevisiae: a yeast homolog of the La autoantigen is dispensable for growth. Mol Cell Biol. 1994 Aug;14(8):5412–5424. doi: 10.1128/mcb.14.8.5412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yoshinaga S. K., Boulanger P. A., Berk A. J. Resolution of human transcription factor TFIIIC into two functional components. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3585–3589. doi: 10.1073/pnas.84.11.3585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Young L. S., Rivier D. H., Sprague K. U. Sequences far downstream from the classical tRNA promoter elements bind RNA polymerase III transcription factors. Mol Cell Biol. 1991 Mar;11(3):1382–1392. doi: 10.1128/mcb.11.3.1382. [DOI] [PMC free article] [PubMed] [Google Scholar]

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