Uncoupling yeast intron recognition from transcription with recursive splicing

Pre‐mRNA splicing has to be coordinated with other processes occurring in the nucleus including transcription, mRNA 3′ end formation and mRNA export. To analyze the relationship between transcription and splicing, we constructed a network of nested introns. Introns were inserted in the 5′ splice site and/or branchpoint of a synthetic yeast intron interrupting a reporter gene. The inserted introns mask the recipient intron from the cellular machinery until they are removed by splicing. Production of functional mRNA from these constructs therefore requires recognition of a spliced RNA as a splicing substrate. We show that recurrent splicing occurs in a sequential and ordered fashion in vivo. Thus, in Saccharomyces cerevisiae, intron recognition and pre‐spliceosome assembly is not tightly coupled to transcription.

[1]  J. Manley,et al.  RNA polymerase II and the integration of nuclear events. , 2000, Genes & development.

[2]  B. Séraphin,et al.  Genomic-scale quantitative analysis of yeast pre-mRNA splicing: implications for splice-site recognition. , 1999, RNA.

[3]  B. Séraphin,et al.  A conditional U5 snRNA mutation affecting pre-mRNA splicing and nuclear pre-mRNA retention identifies SSD1/SRK1 as a general splicing mutant suppressor. , 1999, Nucleic acids research.

[4]  A. Kornblihtt,et al.  Coupling of transcription with alternative splicing: RNA pol II promoters modulate SF2/ASF and 9G8 effects on an exonic splicing enhancer. , 1999, Molecular cell.

[5]  B. Daneholt.  Pre-mRNP particles: From gene to nuclear pore , 1999, Current Biology.

[6]  D. Bentley,et al.  Coupling RNA polymerase II transcription with pre-mRNA processing. , 1999, Current opinion in cell biology.

[7]  A. Lamond,et al.  Nuclear organization of pre-mRNA splicing factors. , 1999, Current Opinion in Cell Biology.

[8]  L. Minvielle-Sebastia,et al.  mRNA polyadenylation and its coupling to other RNA processing reactions and to transcription. , 1999, Current opinion in cell biology.

[9]  J. Manley,et al.  Phosphorylated RNA polymerase II stimulates pre-mRNA splicing. , 1999, Genes & development.

[10]  M. Lai,et al.  A Human Papillomavirus E2 Transcriptional Activator , 1999, The Journal of Biological Chemistry.

[11]  H. Nielsen,et al.  In vivo expression of the nucleolar group I intron‐encoded I‐DirI homing endonuclease involves the removal of a spliceosomal intron , 1999, The EMBO journal.

[12]  D Haussler,et al.  Genome-wide bioinformatic and molecular analysis of introns in Saccharomyces cerevisiae. , 1999, RNA.

[13]  A. J. Lopez,et al.  Generation of alternative Ultrabithorax isoforms and stepwise removal of a large intron by resplicing at exon-exon junctions. , 1998, Molecular cell.

[14]  Han-kuei Huang,et al.  RNA Polymerase I-Promoted HIS4Expression Yields Uncapped, Polyadenylated mRNA That Is Unstable and Inefficiently Translated in Saccharomyces cerevisiae , 1998, Molecular and Cellular Biology.

[15]  M. Roth,et al.  Transcription units as RNA processing units. , 1997, Genes & development.

[16]  M. Ares,et al.  Intron self-complementarity enforces exon inclusion in a yeast pre-mRNA. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[17]  M. Wickens,et al.  The C-terminal domain of RNA polymerase II couples mRNA processing to transcription , 1997, Nature.

[18]  L. Du,et al.  A Functional Interaction between the Carboxy-Terminal Domain of RNA Polymerase II and Pre-mRNA Splicing , 1997, The Journal of cell biology.

[19]  M. Rosbash,et al.  Yeast pre-mRNA is composed of two populations with distinct kinetic properties. , 1996, Experimental cell research.

[20]  C. McGuigan,et al.  A nuclear cap-binding complex facilitates association of U1 snRNP with the cap-proximal 5' splice site. , 1996, Genes & development.

[21]  M. Mathews,et al.  Functional mRNA can be generated by RNA polymerase III , 1995, Molecular and cellular biology.

[22]  H. Klamut,et al.  The human dystrophin gene requires 16 hours to be transcribed and is cotranscriptionally spliced , 1995, Nature Genetics.

[23]  Joe D. Lewis,et al.  A nuclear cap binding protein complex involved in pre-mRNA splicing , 1994, Cell.

[24]  L. Wieslander,et al.  Splicing of Balbiani ring 1 gene pre-mRNA occurs simultaneously with transcription , 1994, Cell.

[25]  A. Greenleaf,et al.  Positive patches and negative noodles: linking RNA processing to transcription? , 1993, Trends in biochemical sciences.

[26]  M. Rosbash,et al.  A yeast splicing factor is localized in discrete subnuclear domains. , 1992, The EMBO journal.

[27]  R. Hallick,et al.  Group II twintron: an intron within an intron in a chloroplast cytochrome b‐559 gene. , 1991, The EMBO journal.

[28]  S. Berget,et al.  Effect of 5' splice site mutations on splicing of the preceding intron , 1990, Molecular and cellular biology.

[29]  C. Thummel,et al.  Splicing precedes polyadenylation during Drosophila E74A transcription , 1990, Molecular and cellular biology.

[30]  K. Köhrer,et al.  A yeast tRNA precursor containing a pre‐mRNA intron is spliced via the pre‐mRNA splicing mechanism. , 1990, The EMBO journal.

[31]  R. Young,et al.  Intragenic and extragenic suppressors of mutations in the heptapeptide repeat domain of Saccharomyces cerevisiae RNA polymerase II. , 1989, Genetics.

[32]  B. Séraphin,et al.  A U1 snRNA:pre‐mRNA base pairing interaction is required early in yeast spliceosome assembly but does not uniquely define the 5′ cleavage site. , 1988, The EMBO journal.

[33]  G. Christofori,et al.  Two spliceosomes can form simultaneously and independently on synthetic double‐intron messenger RNA precursors. , 1987, The EMBO journal.

[34]  M. Rosbash,et al.  mRNA splicing efficiency in yeast and the contribution of nonconserved sequences , 1985, Cell.

[35]  A. M. Miller.  The yeast MATa1 gene contains two introns. , 1984, EMBO Journal.

[36]  L. Guarente,et al.  A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. , 1982, Proceedings of the National Academy of Sciences of the United States of America.