Abstract
In eukarya, RNA is transcribed from a DNA template in the nucleus. The heteronuclear RNA (hnRNA) undergoes further processing, often while being transcribed, to be converted into a messenger RNA (mRNA). The spliceosome acts on hnRNA to remove intervening regions (introns) and join expression regions (exons) using two transesterification reactions. The spliceosome is a large RNA–protein complex composed with over 200 different proteins and five small nuclear RNA (snRNA) that assembles anew on each hnRNA. In this chapter, we will discuss U2-dependent spliceosomal assembly and splicing. We will examine the many steps involved in constitutive splicing using the available cryo-EM structures and associated modeling. The spliceosome is a ribozyme as U6 snRNA and magnesium ions participate in the catalysis reaction.
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References
Harris H, Watts JW. The relationship between nuclear and cytoplasmic ribonucleic acid. Proc R Soc Lond B Biol Sci. 1962; 156:109 –12.
Berget SM, Moore C, Sharp PA. Spliced segments at the 5′ terminus of adenovirus 2 late mRNA. Proc Natl Acad Sci U S A. 1977; 74:3171 –5.
Berk AJ, Sharp PA. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977; 12:721 –32.
Chow LT, Gelinas RE, Broker TR, Roberts RJ. An amazing sequence arrangement at the 5′ ends of adenovirus 2 messenger RNA. Cell. 1977; 12:1 –8.
Darnell JE Jr. Implications of RNA–RNA splicing in evolution of eukaryotic cells. Science. 1978; 202:1257 –60.
Early P, Rogers J, Davis M, et al. Two mRNAs can be produced from a single immunoglobulin chain by alternative RNA processing pathways. Cell. 1980; 20:313 –9.
Berk AJ. Discovery of RNA splicing and genes in pieces. Proc Natl Acad Sci. 2016; 113:801 –5.
Bass B, Cech T. Specific interaction between the self-splicing RNA of Tetrahymena and its guanosine substrate: implications for biological catalysis by RNA. Nature. 1984; 308:820 –6.
Cate JH, Gooding AR, Podell E, et al. Crystal structure of a group I ribozyme domain: principles of RNA packing. Science. 1996; 273:1678 –85.
Su Z, Zhang K, Kappel K, et al. Cryo-EM structures of full-length Tetrahymena ribozyme at 3.1 Å resolution. Nature. 2021; 596:603 –7.
Kashyap L, Sharma RK. Alternative splicing: a paradoxical qudo in eukaryotic genomes. Bioinformation. 2007; 2:155 –6.
Rotival M, Quach H, Quintana-Murci L. Defining the genetic and evolutionary architecture of alternative splicing in response to infection. Nat Commun. 2019; 10:1671 .
Celotto AM, Graveley BR. Alternative splicing of the drosophila Dscam pre-mRNA is both temporally and spatially regulated. Genetics. 2001; 159:599 –608.
Savarese M, Jonson PH, Huovinen S, et al. The complexity of titin splicing pattern in human adult skeletal muscles. Skelet Muscle. 2018; 8:11 .
Will CL, Lührmann R. Spliceosome structure and function. Cold Spring Harb Perspect Biol. 2011;3:a003707.
Guthrie C. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein. Science. 1991; 253:157 –63.
Brow DA, Guthrie C. Spliceosomal RNA U6 is remarkably conserved from yeast to mammals. Nature. 1988; 334:213 –8.
Lee Y, Rio DC. Mechanisms and regulation of alternative pre-mRNA splicing. Annu Rev Biochem. 2015; 84:291 –323.
Bindereif A, Green MR. An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly. EMBO J. 1987; 6:2415 –24.
Feltz C, Anthony K, Brilot A, Pomeranz Krummel D. Architecture of the spliceosome. Biochemistry. 2012; 51:3321 –33.
Raghunathan PL, Guthrie C. RNA unwinding in the U4/U6 snRNPs requires ATP hydrolysis and the DEIH-box splicing factor Brr2. Curr Biol. 1998; 8:847 –55.
Schwer B. A conformational rearrangement in the spliceosome sets the stage for Prp22-dependent mRNA release. Mol Cell. 2008; 30:743 –54.
Weber S, Aebi M. In vitro splicing of mRNA precursors: 5′ cleavage site can be predicted from the interaction between the 5′ splice region and the 5′ terminus of U1 snRNA. Nucleic Acids Res. 1998; 16:471 –86.
Wu J, Manley JL. Mammalian pre-mRNA branch site selection by U2 snRNP involves base pairing. Genes Dev. 1989; 3:1553 –61.
Shi Y. Mechanistic insights into precursor messenger RNA splicing by the spliceosome. Nat Rev Mol Cell Biol. 2017; 18:655 –70.
Cretu C, Gee P, Liu X, et al. Structural basis of intron selection by U2 snRNP in the presence of covalent inhibitors. Nat Commun. 2021; 12:4491 .
Nguyen TH, Galej WP, Bai XC, et al. The architecture of the spliceosomal U4/U6.U5 tri-snRNP. Nature. 2015; 523:47 –52.
Plaschka C, Lin P, Nagai K. Structure of a pre catalytic spliceosome. Nature. 2017; 546:617 –21.
Yan C, Wan R, Bai R, et al. Structure of a yeast activated spliceosome at 3.5 Å resolution. Science. 2016; 353:904 –11.
Galej W, Wilkinson M, Fica S, et al. Cryo-EM structure of the spliceosome immediately after branching. Nature. 2016; 537:197 –201.
Fica SM, Tuttle N, Novak T, et al. RNA catalyses nuclear pre-mRNA splicing. Nature. 2013; 503:229 –34.
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Josefchak, C., Grover, N. (2022). The Spliceosome: A Large Catalytic RNA . In: Grover, N. (eds) Fundamentals of RNA Structure and Function. Learning Materials in Biosciences. Springer, Cham. https://doi.org/10.1007/978-3-030-90214-8_4
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DOI: https://doi.org/10.1007/978-3-030-90214-8_4
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