Abstract
The La autoantigen is an RNA-binding protein that is involved in initiation and termination of RNA polymerase III transcription. It also binds several viral RNAs, including those of poliovirus and human immunodeficiency virus (HIV). Binding of the La protein to these RNAs enhances their translation in vitro (K. Meerovitch, Y.V. Svitkin, H.S. Lee, F. Lejbkowicz, D.J. Kenan, E.K.L. Chan, V.L. Agol, J.D. Keene, and N. Sonenberg, J. Virol. 67:3798-3807, 1993, and Y.V. Svitkin, A. Pause, and N. Sonenberg, J. Virol. 68:7001-7007, 1994). Here, a functional domain in the carboxy-terminal half of La that is distinct from the RNA-binding domain is described. Deletion of this domain abrogated the ability of La protein to enhance translation of poliovirus RNA and a hybrid HIV trans-activation-response element-chloramphenicol acetyltransferase mRNA. Far-Western assays indicated that the La protein homodimerized in vitro, and the C-terminal deletions that caused a loss of activity in translation also abrogated the dimerization signal. Gel filtration chromatography of recombinant La protein confirmed that La protein exists as a dimer under native conditions. Addition of the purified dimerization domain resulted in a loss of translation stimulatory activity of La protein in cell-free-translation reactions.
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- 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]
- Baudin F., Marquet R., Isel C., Darlix J. L., Ehresmann B., Ehresmann C. Functional sites in the 5' region of human immunodeficiency virus type 1 RNA form defined structural domains. J Mol Biol. 1993 Jan 20;229(2):382–397. doi: 10.1006/jmbi.1993.1041. [DOI] [PubMed] [Google Scholar]
- 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]
- Blanar M. A., Rutter W. J. Interaction cloning: identification of a helix-loop-helix zipper protein that interacts with c-Fos. Science. 1992 May 15;256(5059):1014–1018. doi: 10.1126/science.1589769. [DOI] [PubMed] [Google Scholar]
- Casas-Finet J. R., Smith J. D., Jr, Kumar A., Kim J. G., Wilson S. H., Karpel R. L. Mammalian heterogeneous ribonucleoprotein A1 and its constituent domains. Nucleic acid interaction, structural stability and self-association. J Mol Biol. 1993 Feb 20;229(4):873–889. doi: 10.1006/jmbi.1993.1093. [DOI] [PubMed] [Google Scholar]
- 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]
- Chan E. K., Tan E. M. Epitopic targets for autoantibodies in systemic lupus erythematosus and Sjögren's syndrome. Curr Opin Rheumatol. 1989 Oct;1(3):376–381. doi: 10.1097/00002281-198901030-00022. [DOI] [PubMed] [Google Scholar]
- 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]
- Dorner A. J., Semler B. L., Jackson R. J., Hanecak R., Duprey E., Wimmer E. In vitro translation of poliovirus RNA: utilization of internal initiation sites in reticulocyte lysate. J Virol. 1984 May;50(2):507–514. doi: 10.1128/jvi.50.2.507-514.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edery I., Petryshyn R., Sonenberg N. Activation of double-stranded RNA-dependent kinase (dsl) by the TAR region of HIV-1 mRNA: a novel translational control mechanism. Cell. 1989 Jan 27;56(2):303–312. doi: 10.1016/0092-8674(89)90904-5. [DOI] [PubMed] [Google Scholar]
- Evdokimova V. M., Wei C. L., Sitikov A. S., Simonenko P. N., Lazarev O. A., Vasilenko K. S., Ustinov V. A., Hershey J. W., Ovchinnikov L. P. The major protein of messenger ribonucleoprotein particles in somatic cells is a member of the Y-box binding transcription factor family. J Biol Chem. 1995 Feb 17;270(7):3186–3192. doi: 10.1074/jbc.270.7.3186. [DOI] [PubMed] [Google Scholar]
- Fresco L. D., Harper D. S., Keene J. D. Leucine periodicity of U2 small nuclear ribonucleoprotein particle (snRNP) A' protein is implicated in snRNP assembly via protein-protein interactions. Mol Cell Biol. 1991 Mar;11(3):1578–1589. doi: 10.1128/mcb.11.3.1578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geballe A. P., Gray M. K. Variable inhibition of cell-free translation by HIV-1 transcript leader sequences. Nucleic Acids Res. 1992 Aug 25;20(16):4291–4297. doi: 10.1093/nar/20.16.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Hermann H., Fabrizio P., Raker V. A., Foulaki K., Hornig H., Brahms H., Lührmann R. snRNP Sm proteins share two evolutionarily conserved sequence motifs which are involved in Sm protein-protein interactions. EMBO J. 1995 May 1;14(9):2076–2088. doi: 10.1002/j.1460-2075.1995.tb07199.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herschlag D. RNA chaperones and the RNA folding problem. J Biol Chem. 1995 Sep 8;270(36):20871–20874. doi: 10.1074/jbc.270.36.20871. [DOI] [PubMed] [Google Scholar]
- Jones K. A., Peterlin B. M. Control of RNA initiation and elongation at the HIV-1 promoter. Annu Rev Biochem. 1994;63:717–743. doi: 10.1146/annurev.bi.63.070194.003441. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Marello K., LaRovere J., Sommerville J. Binding of Xenopus oocyte masking proteins to mRNA sequences. Nucleic Acids Res. 1992 Nov 11;20(21):5593–5600. doi: 10.1093/nar/20.21.5593. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meerovitch K., Pelletier J., Sonenberg N. A cellular protein that binds to the 5'-noncoding region of poliovirus RNA: implications for internal translation initiation. Genes Dev. 1989 Jul;3(7):1026–1034. doi: 10.1101/gad.3.7.1026. [DOI] [PubMed] [Google Scholar]
- 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]
- Miele G., Mouland A., Harrison G. P., Cohen E., Lever A. M. The human immunodeficiency virus type 1 5' packaging signal structure affects translation but does not function as an internal ribosome entry site structure. J Virol. 1996 Feb;70(2):944–951. doi: 10.1128/jvi.70.2.944-951.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Méthot N., Song M. S., Sonenberg N. A region rich in aspartic acid, arginine, tyrosine, and glycine (DRYG) mediates eukaryotic initiation factor 4B (eIF4B) self-association and interaction with eIF3. Mol Cell Biol. 1996 Oct;16(10):5328–5334. doi: 10.1128/mcb.16.10.5328. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardigon N., Strauss J. H. Mosquito homolog of the La autoantigen binds to Sindbis virus RNA. J Virol. 1996 Feb;70(2):1173–1181. doi: 10.1128/jvi.70.2.1173-1181.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parkin N. T., Cohen E. A., Darveau A., Rosen C., Haseltine W., Sonenberg N. Mutational analysis of the 5' non-coding region of human immunodeficiency virus type 1: effects of secondary structure on translation. EMBO J. 1988 Sep;7(9):2831–2837. doi: 10.1002/j.1460-2075.1988.tb03139.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pelletier J., Sonenberg N. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature. 1988 Jul 28;334(6180):320–325. doi: 10.1038/334320a0. [DOI] [PubMed] [Google Scholar]
- SenGupta D. N., Berkhout B., Gatignol A., Zhou A. M., Silverman R. H. Direct evidence for translational regulation by leader RNA and Tat protein of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7492–7496. doi: 10.1073/pnas.87.19.7492. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Svitkin Y. V., Maslova S. V., Agol V. I. The genomes of attenuated and virulent poliovirus strains differ in their in vitro translation efficiencies. Virology. 1985 Dec;147(2):243–252. doi: 10.1016/0042-6822(85)90127-8. [DOI] [PubMed] [Google Scholar]
- Svitkin Y. V., Meerovitch K., Lee H. S., Dholakia J. N., Kenan D. J., Agol V. I., Sonenberg N. Internal translation initiation on poliovirus RNA: further characterization of La function in poliovirus translation in vitro. J Virol. 1994 Mar;68(3):1544–1550. doi: 10.1128/jvi.68.3.1544-1550.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Svitkin Y. V., Pause A., Sonenberg N. La autoantigen alleviates translational repression by the 5' leader sequence of the human immunodeficiency virus type 1 mRNA. J Virol. 1994 Nov;68(11):7001–7007. doi: 10.1128/jvi.68.11.7001-7007.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tanchou V., Gabus C., Rogemond V., Darlix J. L. Formation of stable and functional HIV-1 nucleoprotein complexes in vitro. J Mol Biol. 1995 Oct 6;252(5):563–571. doi: 10.1006/jmbi.1995.0520. [DOI] [PubMed] [Google Scholar]