Recognition of the 5' splice site by the spliceosome

• Autorzy: Konarska M. M.
• Języki publikacji: EN

Abstrakty

EN

The splicing of nuclear pre-mRNAs is catalyzed by a large, multicomponent ribonucleoprotein complex termed the spliceosome. Elucidation of the molecular mechanism of splicing identified small nuclear RNAs (snRNAs) as important components of the spliceosome, which, by analogy to the self-splicing group II introns, are implicated in formation of the catalytic center. In particular, the 5' splice site (5'SS) and the branch site, which represent the two substrates for the first step of splicing, are first recognized by U1 and U2 snRNPs, respectively. This initial recognition of splice sites is responsible for the global definition of exons and introns, and represents the primary target for regulation of splicing. Subsequently, pairing interaction between the 5'SS and U1 snRNA is disrupted and replaced by a new interaction of the 5'SS with U6 snRNA. The 5'SS signal contains an invariant GU dinucleotide present at the 5' end of nearly all known introns, however, the mechanism by which the spliceosome recognizes this element is not known. We have identified and characterized a specific UV light-induced crosslink formed between the 5'SS RNA and hPrp8, a protein component of U5 snRNP in the spliceosome that is likely to reflect a specific recognition of the GU dinucleotide for splicing. Because recognition of the 5'SS must be linked to formation of the catalytic site, the identification of a specific and direct interaction between the 5'SS and Prp8 has significant implications for the role of this protein in the mechanism of mRNA splicing.

Słowa kluczowe

EN

mRNA splicing; pre-mRNA splicing; protein; splicing; spliceosome; ribonucleoprotein complex; splice site

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1998
• Tom: 45
• Numer: 4
• Opis fizyczny: p.869-881,fig.

Twórcy

autor

Konarska M. M.

• The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA

Bibliografia

• 1. Kramer, A. (1996) The structure and function of proteins involved in mammalian pre-mRNA splicing. Annu. Rev. Biochem. 65, 367-409.
• 2. Moore, M.J., Query, C.C. & Sharp, P.A. (1993) Splicing of precursors to mRNA by the spli­ceosome; in The RNA World (Gesteland, R.F & Atkins, J.F., eds.) pp. 303-357, Cold Spring Harbor Laboratory Press, Cold Spring Har­bor, New York.
• 3 Nilsen, T.W. (1998) RNA-RNA interactions in nuclear pre-mRNA splicing; in RNA Structure and Function (Simons, R.W. & Grunberg- Manago, M.f eds.) pp. 279-307, Cold Spring Harbor Laboratory Press, New York.
• 4. Nilsen, T.W. (1994) RNA-RNA interactions in the spliceosome: Unraveling the ties that bind. Cell 78, 1-4.
• 5. Staley, J.P. & Guthrie, C. (1998) Mechanical devices of the spliceosome: Motors, clocks, springs, and things. Cell 92, 315-326.
• 6. Hertel, K.J., Lynch, K.W. & Maniatis, T. (1997) Common themes in the function of transcription and splicing enhancers. Curr. Opin. Cell Biol 9, 350-357.
• 7. Newman, A. (1997) RNA splicing: Out of the loop. Curr. Biol. 7, 418-420.
• 8. Wang, J. & Manley, J.L. (1997) Regulation of pre-mRNA splicing in metazoa. Curr. Opin. Genet. Dev. 7, 205-211.
• 9. Will, C.L. à Luhrmann, R. (1997) Protein functions in pre-mRNA splicing. Curr. Opin. Cell Biol. 9, 320-328.
• 10. Reed, R. (1996) Initial splice-site recognition and pairing during pre-mRNA splicing. Curr. Opin. GeneL Dev. 6, 215-220.
• 11. Guthrie, C. (1994) The spliceosome is a dy­namic ribonucleoprotein machine. Harvey Lect. 90, 59-80.
• 12.0'Keefe, R.T., Norman, C. & Newman, A.J. (1996) The invariant U5 snRNA loop 1 se­quence is dispensable for the first catalytic step of pre-mRNA splicing in yeast. Cell 86. 679-689.
• 13.0'Keefe, R.T. & Newman, A.J. (1998) Func­tional analysis of the U5 snRNA loop 1 in the second catalytic step of yeast pre-mRNA splic­ing. EMBO J. 17, 565-574.
• 14. Hall, S.L. & Padgett, R.A. (1994) Conserved sequences in a class of rare eukaryotic nuclear introns with non-consensus splice sites. J. Mol Biol 239. 357-365.
• 15. Hall, S.L. & Padgett, R.A. (1996) Requirement of U12 snRNA for in vivo splicing of a minor class of eukaryotic nuclear pre-mRNA introns (see comments). Science 271, 1716-1718.
• 16. Tarn, W.-Y. & Steitz, J.A. (1996) A novel spli­ceosome containing Ull, U12, and U5 snRNPs excises a minor class (AT-AC) intron in vitro. Cell 84. 801-811.
• 17. Tarn, W.Y. & Steitz, J.A. (1996) Highly di­verged U4 and U6 small nuclear RNAs re­quired for splicing rare AT-AC introns (see comments). Science 273, 1824-1832.
• 18. Incorvaia, K. & Padgett, R.A. (1998) Base pair­ing with U6atac is required for 5' splice site activation of Ul2-dependent introns in vivo. RNA 4, 709-718.
• 19. Cech, T.R. (1983) RNA splicing: Three themes with variations. Cell 34, 713-716.
• 20. Sharp, P.A. (1991) "Five easy pieces'. Science 254, 663.
• 21. Sharp, P.A. (1994) Split genes and RNA splic­ing. CeU 77, 805-815.
• 22. Guthrie, C. (1991) Messenger RNA splicing in yeast: Clues to why the spliceosome is a ribo­nucleoprotein. Science 253, 157-163.
• 23. Crispino, J.D., Blencowe, B.J. & Sharp, P.A. (1994) Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP. Science 265. 1866-1869.
• 24. Tarn, W.-Y. & Steitz, J.A. (1994) SR proteins can compensate for the loss of U1 snRNP functions in vitro. Genes & Dev. 8,2704-2717.
• 25. Robberson, B.L., Cote, GJ. & Berget, S.M. (1990) Exon definition may facilitate splice site selection in RNAs with multiple exons. Mol. Cell Biol 10, 84-94.
• 26. Berget, S.M. (1995) Exon recognition in verte­brate splicing. J. Biol Chem. 270, 2411- 2414.
• 27. Abovich, N.. Liao, X.C. & Rosbash, M. (1994) The yeast MUD2 protein: An interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition. Genes & Dev. 8, 843-854.
• 28. Legrain, P., Seraphin, B. & Rosbash, M. (1988) Early commitment of yeast pre-mRNA to the spliceosome pathway. Mol Cell. Biol 8, 3755-3760.
• 29.Staknis, D. & Reed, R. (1994) SR proteins pro­mote the first specific recognition of pre- mRNA and are present together with U1 small ribonucleoprotein in a general splicing enhan­cer complex. Mol Cell Biol 14, 7670-7682.
• 30. Crispino, J.D., Mermoud, J.E., Lamond, A.I. & Sharp, P.A. (1996) Cis-acting elements dis­tinct from the 5' splice site promote U1- independent pre-mRNA splicing. RNA 2, 664-673.
• 31. Alvarez, C.J., Romfo, C.M., Vanhoy, R.W., Porter, G.L. & Wise, J.A. (1996) Mutational analysis of U1 function in Schizosaccharomy- ces pombe: pre-mRNAs differ in the extent and nature of their requirements for this snRNA in vivo. RNA 2, 404-418.
• 32. Konforti, B.B. & Konarska, M.M. (1995) A short 5' splice site RNA oligo can participate in both steps of splicing in mammalian ex­tracts. RNA 1, 815-827.
• 33. Konforti, B.B., Koziolkiewicz. M.J. & Konar­ska, M.M. (1993) Disruption of base pairing between the 5' splice site and the 5' end of U1 snRNA is required for spliceosome assembly. Cell 75, 863-873.
• 34. Hall, K.B. & Konarska, M.M. (1992) The 5' splice site consensus RNA oligonucleotide in­duces assembly of U2/U4/U5/U6 small nu­clear ribonucleoprotein complexes. Proc. NatL Acad. Sci. U.S.A. 89, 10969-10973.
• 35. Maroney, P.A., Hannon, G.J., Shambaugh, J.D. & Nilsen, T.W. (1991) Intramolecular base pairing between the nematode spliced leader and its 5' splice site is not essential for irans-splicing in vitro. EMBO J. 10, 3869- 3875.
• 36. Bruzik, J.P. & Steitz, J.A. (1990) Spliced leader RNA sequences can substitute for the essential 5' end of U1 RNA during splicing in a mammalian in vitro system. Cell 62, 889- 899.
• 37. Madhani, H.D. & Guthrie, C. (1994) Dynamic RNA-RNA interactions in the spliceosome. Annu. Rev. Genet 28, 1-26.
• 38. Reyes, J.L., Kois, P., Konforti, B.B. & Konar­ska, M.M. (1996) The canonical GU dinucleo- tide at the 5' splice site is recognized by p220 of the U5 snRNP within the spliceosome. RNA 2, 213-225.
• 39. Reyes, J.L. (1998) Biochemical and genetic analysis of the interaction of Prp8 with the 5' splice site during pre-mRNA splicing; in Labo­ratory of Molecular Biology and Biochemistry. The Rockefeller University, New York.
• 40. Crispino, J.D. & Sharp, P.A. (1995) A U6 snRNA:pre-mRNA interaction can be rate-limiting for Ul-independent splicing. Genes & Dev. 9, 2314-2323.
• 41. Dietrich, R.C., Incorvaia, R. & Padgett, R.A. (1997) Terminal intron dinucleotide se­quences do not distinguish between U2- and U12-dependent introns. Moi Cell 1,151-160.
• 42.Sharp, P.A. & Bürge, C.B. (1997) Classifica­tion of introns: U2-type or U12-type. Cell 91, 875-879.
• 43. Hodges, P.E., Jackson, S.P., Brown, J.D. & Beggs, J.D. (1995) Extraordinary sequence conservation of the PRP8 splicing factor. Yeast 11, 337-342.
• 44. Lücke, S., Klöckner, T., Palfi, Z., Boshart, M. & Binderief, A. (1997) Trans mRNA splicing in trypanosomes: Cloning and analysis of a P/?P
• 45. Wyatt, J.R., Sontheimer, E.J. & Steitz. J.A. (1992) Site-specific cross-linking of mammal­ian U5 snRNP to the 5' splice site before the first step of pre-mRNA splicing. Genes & Dev. 6, 2542-2553.
• 46. Teigelkamp, S., Newman, A.J. & Beggs, J.D. (1995) Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon align­ment by U5 snRNA. EMBO J. 14, 2602-2612.
• 47. Chiara, M.D., Gozani, O., Bennett, M., Cham­pion-Arnaud, P., Palandjian, L. & Reed, R. (1996) Identification of proteins that interact with exon sequences, splice sites, and the branchpoint sequence during each stage of spliceosome assembly. MoL Cell. Biol. 16, 3317-3326.
• 48. MacMillan, A.M., Query, Ch.C., Allerson, Ch.R., Chen, S., Verdine, G.L. & Sharp, P.A. (1994) Dynamic association of proteins with the pre-mRNA branch region. Genes & Dev. 8, 3008-3020.
• 49. Umen, J.G. & Guthrie, C. (1995) A novel role for a U5 snRNP protein in 3' splice site selec­tion. Genes & Dev. 9, 855-868.
• 50. Achsel, T., Ahrens, K., Brahms, H., Teigel- kamp, S. & Lührmann, R. (1998) The human U5-220kD protein (hPrp8) forms a stable RNA-free complex with several U5-specific proteins, including an RNA unwindase, a homologue of ribosomal elongation factor EF- 2, and a novel WEMO protein. Mol Cell Biol 18, 6756-6766.
• 51. Dix, I., Rüssel, C.S., O'Keefe, R.T., Newman, A.J. & Beggs. J.D. (1998) Protein-RNA inter­actions in the U5 snRNP of Saccharomyces cerevisiae. RNA 4, 1239-1250.
• 52. Teigelkamp, S.. Mundt. C., Achsel. T.. Will, C.L. & Luhrmann, R. (1997) The human U5 snRNP-specific 100-kD protein is an RS domain-containing, putative RNA helicase with significant homology to the yeast splicing factor Prp28p. RNA 3, 1313-1326.
• 53. Reyes, J.L., Gustafson, E.H., Luo, H.R., Moore, M.J. & Konarska, M.M. (1998) The C- terminal region of hPrp8 interacts with the conserved GU dinucleotide at the 5' splice site. RNA, in press.