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. 1991 Oct 11;19(19):5213–5217. doi: 10.1093/nar/19.19.5213

Evolutionary conservation of the spliceosomal protein, U2B''.

G G Simpson 1, P Vaux 1, G Clark 1, R Waugh 1, J D Beggs 1, J W Brown 1
PMCID: PMC328878  PMID: 1833724

Abstract

U1 and U2snRNPs play key roles in pre-mRNA splicing. The interactions between the U1 and U2snRNP-specific proteins, U1A, U2A' and U2B'' and their respective UsnRNAs are of interest both to elucidate their roles in splicing, and as models to study RNA-protein interactions. We have cloned a full-length cDNA, encoding U2B'', from potato. This is the first report of a sequence for a plant UsnRNP protein. The plant U2B'' sequence exhibits extensive similarity with the human U2B'' protein at both the DNA and amino acid levels. The evolutionary conservation at the protein level, particularly in sequences implicated in determining specific binding to U2snRNA, suggests conservation of U2B'' function from plants to man. The significance of amino acid substitutions in the RNP-80 motif with respect to U2snRNA binding in plants is discussed.

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Selected References

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  1. Abel S., Kiss T., Solymosy F. Molecular analysis of eight U1 RNA gene candidates from tomato that could potentially be transcribed into U1 RNA sequence variants differing from each other in similar regions of secondary structure. Nucleic Acids Res. 1989 Aug 11;17(15):6319–6337. doi: 10.1093/nar/17.15.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adam S. A., Nakagawa T., Swanson M. S., Woodruff T. K., Dreyfuss G. mRNA polyadenylate-binding protein: gene isolation and sequencing and identification of a ribonucleoprotein consensus sequence. Mol Cell Biol. 1986 Aug;6(8):2932–2943. doi: 10.1128/mcb.6.8.2932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Boelens W., Scherly D., Beijer R. P., Jansen E. J., Dathan N. A., Mattaj I. W., van Venrooij W. J. A weak interaction between the U2A' protein and U2 snRNA helps to stabilize their complex with the U2B" protein. Nucleic Acids Res. 1991 Feb 11;19(3):455–460. doi: 10.1093/nar/19.3.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown J. W., Waugh R. Maize U2 snRNAs: gene sequence and expression. Nucleic Acids Res. 1989 Nov 25;17(22):8991–9001. doi: 10.1093/nar/17.22.8991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fritz A., Parisot R., Newmeyer D., De Robertis E. M. Small nuclear U-ribonucleoproteins in Xenopus laevis development. Uncoupled accumulation of the protein and RNA components. J Mol Biol. 1984 Sep 15;178(2):273–285. doi: 10.1016/0022-2836(84)90144-x. [DOI] [PubMed] [Google Scholar]
  8. Gómez J., Sánchez-Martínez D., Stiefel V., Rigau J., Puigdomènech P., Pagès M. A gene induced by the plant hormone abscisic acid in response to water stress encodes a glycine-rich protein. Nature. 1988 Jul 21;334(6179):262–264. doi: 10.1038/334262a0. [DOI] [PubMed] [Google Scholar]
  9. Habets W. J., Hoet M. H., De Jong B. A., Van der Kemp A., Van Venrooij W. J. Mapping of B cell epitopes on small nuclear ribonucleoproteins that react with human autoantibodies as well as with experimentally-induced mouse monoclonal antibodies. J Immunol. 1989 Oct 15;143(8):2560–2566. [PubMed] [Google Scholar]
  10. Habets W. J., Sillekens P. T., Hoet M. H., Schalken J. A., Roebroek A. J., Leunissen J. A., van de Ven W. J., van Venrooij W. J. Analysis of a cDNA clone expressing a human autoimmune antigen: full-length sequence of the U2 small nuclear RNA-associated B" antigen. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2421–2425. doi: 10.1073/pnas.84.8.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Habets W., Hoet M., Bringmann P., Lührmann R., van Venrooij W. Autoantibodies to ribonucleoprotein particles containing U2 small nuclear RNA. EMBO J. 1985 Jun;4(6):1545–1550. doi: 10.1002/j.1460-2075.1985.tb03815.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hamm J., Dathan N. A., Mattaj I. W. Functional analysis of mutant Xenopus U2 snRNAs. Cell. 1989 Oct 6;59(1):159–169. doi: 10.1016/0092-8674(89)90878-7. [DOI] [PubMed] [Google Scholar]
  13. Hanley B. A., Schuler M. A. Nucleotide sequence of a pea U2 snRNA gene. Nucleic Acids Res. 1989 Dec 11;17(23):10106–10106. doi: 10.1093/nar/17.23.10106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hanley B. A., Schuler M. A. cDNA cloning of U1, U2, U4 and U5 snRNA families expressed in pea nuclei. Nucleic Acids Res. 1991 Apr 25;19(8):1861–1869. doi: 10.1093/nar/19.8.1861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Kiss T., Solymosy F. Molecular analysis of a U3 RNA gene locus in tomato: transcription signals, the coding region, expression in transgenic tobacco plants and tandemly repeated pseudogenes. Nucleic Acids Res. 1990 Apr 25;18(8):1941–1949. doi: 10.1093/nar/18.8.1941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lehmeier T., Foulaki K., Lührmann R. Evidence for three distinct D proteins, which react differentially with anti-Sm autoantibodies, in the cores of the major snRNPs U1, U2, U4/U6 and U5. Nucleic Acids Res. 1990 Nov 25;18(22):6475–6484. doi: 10.1093/nar/18.22.6475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Li Y. Q., Sugiura M. Three distinct ribonucleoproteins from tobacco chloroplasts: each contains a unique amino terminal acidic domain and two ribonucleoprotein consensus motifs. EMBO J. 1990 Oct;9(10):3059–3066. doi: 10.1002/j.1460-2075.1990.tb07502.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Li Y. Q., Ye L. Z., Sugita M., Sugiura M. Tobacco nuclear gene for the 31 kd chloroplast ribonucleoprotein: genomic organization, sequence analysis and expression. Nucleic Acids Res. 1991 Jun 11;19(11):2987–2991. doi: 10.1093/nar/19.11.2987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lührmann R., Kastner B., Bach M. Structure of spliceosomal snRNPs and their role in pre-mRNA splicing. Biochim Biophys Acta. 1990 Nov 30;1087(3):265–292. doi: 10.1016/0167-4781(90)90001-i. [DOI] [PubMed] [Google Scholar]
  21. Mancebo R., Lo P. C., Mount S. M. Structure and expression of the Drosophila melanogaster gene for the U1 small nuclear ribonucleoprotein particle 70K protein. Mol Cell Biol. 1990 Jun;10(6):2492–2502. doi: 10.1128/mcb.10.6.2492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Marshallsay C., Kiss T., Filipowicz W. Amplification of plant U3 and U6 snRNA gene sequences using primers specific for an upstream promoter element and conserved intragenic regions. Nucleic Acids Res. 1990 Jun 25;18(12):3459–3466. doi: 10.1093/nar/18.12.3459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Marshallsay C., Kiss T., Filipowicz W. Amplification of plant U3 and U6 snRNA gene sequences using primers specific for an upstream promoter element and conserved intragenic regions. Nucleic Acids Res. 1990 Jun 25;18(12):3459–3466. doi: 10.1093/nar/18.12.3459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mattaj I. W. A binding consensus: RNA-protein interactions in splicing, snRNPs, and sex. Cell. 1989 Apr 7;57(1):1–3. doi: 10.1016/0092-8674(89)90164-5. [DOI] [PubMed] [Google Scholar]
  25. Mattaj I. W., De Robertis E. M. Nuclear segregation of U2 snRNA requires binding of specific snRNP proteins. Cell. 1985 Jan;40(1):111–118. doi: 10.1016/0092-8674(85)90314-9. [DOI] [PubMed] [Google Scholar]
  26. Mortenson E., Dreyfuss G. RNP in maize protein. Nature. 1989 Jan 26;337(6205):312–312. doi: 10.1038/337312b0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Pan Z. Q., Prives C. U2 snRNA sequences that bind U2-specific proteins are dispensable for the function of U2 snRNP in splicing. Genes Dev. 1989 Dec;3(12A):1887–1898. doi: 10.1101/gad.3.12a.1887. [DOI] [PubMed] [Google Scholar]
  29. Pálfi Z., Bach M., Solymosy F., Lührmann R. Purification of the major UsnRNPs from broad bean nuclear extracts and characterization of their protein constituents. Nucleic Acids Res. 1989 Feb 25;17(4):1445–1458. doi: 10.1093/nar/17.4.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Ruby S. W., Abelson J. Pre-mRNA splicing in yeast. Trends Genet. 1991 Mar;7(3):79–85. doi: 10.1016/0168-9525(91)90276-V. [DOI] [PubMed] [Google Scholar]
  32. Sachs A. B., Bond M. W., Kornberg R. D. A single gene from yeast for both nuclear and cytoplasmic polyadenylate-binding proteins: domain structure and expression. Cell. 1986 Jun 20;45(6):827–835. doi: 10.1016/0092-8674(86)90557-x. [DOI] [PubMed] [Google Scholar]
  33. Scherly D., Boelens W., Dathan N. A., van Venrooij W. J., Mattaj I. W. Major determinants of the specificity of interaction between small nuclear ribonucleoproteins U1A and U2B'' and their cognate RNAs. Nature. 1990 Jun 7;345(6275):502–506. doi: 10.1038/345502a0. [DOI] [PubMed] [Google Scholar]
  34. Scherly D., Boelens W., van Venrooij W. J., Dathan N. A., Hamm J., Mattaj I. W. Identification of the RNA binding segment of human U1 A protein and definition of its binding site on U1 snRNA. EMBO J. 1989 Dec 20;8(13):4163–4170. doi: 10.1002/j.1460-2075.1989.tb08601.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Schuster G., Gruissem W. Chloroplast mRNA 3' end processing requires a nuclear-encoded RNA-binding protein. EMBO J. 1991 Jun;10(6):1493–1502. doi: 10.1002/j.1460-2075.1991.tb07669.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shuster E. O., Guthrie C. Two conserved domains of yeast U2 snRNA are separated by 945 nonessential nucleotides. Cell. 1988 Oct 7;55(1):41–48. doi: 10.1016/0092-8674(88)90007-4. [DOI] [PubMed] [Google Scholar]
  38. Sillekens P. T., Beijer R. P., Habets W. J., van Venrooij W. J. Human U1 snRNP-specific C protein: complete cDNA and protein sequence and identification of a multigene family in mammals. Nucleic Acids Res. 1988 Sep 12;16(17):8307–8321. doi: 10.1093/nar/16.17.8307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sillekens P. T., Beijer R. P., Habets W. J., van Verooij W. J. Molecular cloning of the cDNA for the human U2 snRNA-specific A' protein. Nucleic Acids Res. 1989 Mar 11;17(5):1893–1906. doi: 10.1093/nar/17.5.1893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sillekens P. T., Habets W. J., Beijer R. P., van Venrooij W. J. cDNA cloning of the human U1 snRNA-associated A protein: extensive homology between U1 and U2 snRNP-specific proteins. EMBO J. 1987 Dec 1;6(12):3841–3848. doi: 10.1002/j.1460-2075.1987.tb02721.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Szkukálek A., Kiss T., Solymosy F. The 5' end of the coding region of a U6 RNA gene candidate from tomato starts with GUCC, a phylogenetically highly conserved 5' end sequence of U6 RNA. Nucleic Acids Res. 1990 Mar 11;18(5):1295–1295. doi: 10.1093/nar/18.5.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Theissen H., Etzerodt M., Reuter R., Schneider C., Lottspeich F., Argos P., Lührmann R., Philipson L. Cloning of the human cDNA for the U1 RNA-associated 70K protein. EMBO J. 1986 Dec 1;5(12):3209–3217. doi: 10.1002/j.1460-2075.1986.tb04631.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tollervey D., Mattaj I. W. Fungal small nuclear ribonucleoproteins share properties with plant and vertebrate U-snRNPs. EMBO J. 1987 Feb;6(2):469–476. doi: 10.1002/j.1460-2075.1987.tb04777.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Vankan P., Filipowicz W. Structure of U2 snRNA genes of Arabidopsis thaliana and their expression in electroporated plant protoplasts. EMBO J. 1988 Mar;7(3):791–799. doi: 10.1002/j.1460-2075.1988.tb02877.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Waugh R., Clark G., Vaux P., Brown J. W. Sequence and expression of potato U2 snRNA genes. Nucleic Acids Res. 1991 Jan 25;19(2):249–256. doi: 10.1093/nar/19.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. van Santen V. L., Spritz R. A. Nucleotide sequence of a bean (Phaseolus vulgaris) U1 small nuclear RNA gene: implications for plant pre-mRNA splicing. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9094–9098. doi: 10.1073/pnas.84.24.9094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. van Santen V. L., Swain W., Spritz R. A. Nucleotide sequences of two soybean U1 snRNA genes. Nucleic Acids Res. 1988 May 11;16(9):4176–4176. doi: 10.1093/nar/16.9.4176. [DOI] [PMC free article] [PubMed] [Google Scholar]

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