Connections between Pre-mRNA Processing and Regulation of the Eukaryotic Cell Cycle

Burns, Claire; Gould, Kathy

doi:10.1159/000060995

Cited by 10 publications

(6 citation statements)

References 107 publications

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“…Such varied phenotypes can arise from defective pre-mRNA splicing [64] if spliceosome assembly is affected. The predicted consequence of defective spliceosome assembly is that variable mRNAs become limiting at different times, and the cells eventually die from accumulated defects in multiple processes.…”

Section: Discussionmentioning

confidence: 99%

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

et al. 2011

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Prp19 is the founding member of the NineTeen Complex, or NTC, which is a spliceosomal subcomplex essential for spliceosome activation. To define Prp19 connectivity and dynamic protein interactions within the spliceosome, we systematically queried the Saccharomyces cerevisiae proteome for Prp19 WD40 domain interaction partners by two-hybrid analysis. We report that in addition to S. cerevisiae Cwc2, the splicing factor Prp17 binds directly to the Prp19 WD40 domain in a 1∶1 ratio. Prp17 binds simultaneously with Cwc2 indicating that it is part of the core NTC complex. We also find that the previously uncharacterized protein Urn1 (Dre4 in Schizosaccharomyces pombe) directly interacts with Prp19, and that Dre4 is conditionally required for pre-mRNA splicing in S. pombe. S. pombe Dre4 and S. cerevisiae Urn1 co-purify U2, U5, and U6 snRNAs and multiple splicing factors, and dre4Δ and urn1Δ strains display numerous negative genetic interactions with known splicing mutants. The S. pombe Prp19-containing Dre4 complex co-purifies three previously uncharacterized proteins that participate in pre-mRNA splicing, likely before spliceosome activation. Our multi-faceted approach has revealed new low abundance splicing factors connected to NTC function, provides evidence for distinct Prp19 containing complexes, and underscores the role of the Prp19 WD40 domain as a splicing scaffold.

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Section: Discussionmentioning

confidence: 99%

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

et al. 2011

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“…Finally, the cell cycle‐dependent accumulation of phospho‐Thr 199 NPM at nuclear speckles mediated by CDK2/cyclin E (and A) and the involvement of phospho‐Thr 199 NPM in pre‐mRNA splicing process raise the interesting question regarding to a possible link between pre‐mRNA splicing and cell cycle progression. Indeed, there is growing evidence linking these two events (reviewed in [57,58]). For instance, one of SR protein like factor SRp38 (also known as SRrp40, TASR‐2, NSSR‐1) was shown to act as a splicing repressor when dephosphorylated during M phase, and it was suggested that dephosphorylation of SRp38 plays a role in gene silencing during mitosis [59].…”

Section: Discussionmentioning

confidence: 99%

Thr¹⁹⁹ phosphorylation targets nucleophosmin to nuclear speckles and represses pre‐mRNA processing

et al. 2005

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Nucleophosmin (NPM) is a multifunctional phosphoprotein, being involved in ribosome assembly, pre-ribosomal RNA processing, DNA duplication, nucleocytoplasmic protein trafficking, and centrosome duplication. NPM is phosphorylated by several kinases, including nuclear kinase II, casein kinase 2, Polo-like kinase 1 and cyclin-dependent kinases (CDK1 and 2), and these phosphorylations modulate the activity and function of NPM. We have previously identified Thr 199 as the major phosphorylation site of NPM mediated by CDK2/cyclin E (and A), and this phosphorylation is involved in the regulation of centrosome duplication. In this study, we further examined the effect of CDK2-mediated phosphorylation of NPM by using the antibody that specifically recognizes NPM phosphorylated on Thr 199 . We found that the phospho-Thr 199 NPM localized to dynamic sub-nuclear structures known as nuclear speckles, which are believed to be the sites of storage and/or assembly of premRNA splicing factors. Phosphorylation on Thr 199 by CDK2/cyclin E (and A) targets NPM to nuclear speckles, and enhances the RNA-binding activity of NPM. Moreover, phospho-Thr 199 NPM, but not unphosphorylated NPM, effectively represses pre-mRNA splicing. These findings indicate the involvement of NPM in the regulation of pre-mRNA processing, and its activity is controlled by CDK2-mediated phosphorylation on Thr 199 .

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“…Substantial evidence has accumulated to suggest that these Myb-related proteins play an essential role in RNA processing rather than transcription (3,10,69). Genetic depletion of S. pombe Cdc5p or S. cerevisiae Cef1p causes accumulation of unspliced mRNAs in vivo (9,43,69). Inactivation of Cef1p by antibody interference or immunodepletion of hCDC5 inhibits splicing in vitro (3, 69).…”

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confidence: 99%

Proteomics Analysis Reveals Stable Multiprotein Complexes in Both Fission and Budding Yeasts Containing Myb-Related Cdc5p/Cef1p, Novel Pre-mRNA Splicing Factors, and snRNAs

Ohi

Link

Ren

et al. 2002

Molecular and Cellular Biology

202

217

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Schizosaccharomyces pombe Cdc5p and its Saccharomyces cerevisiae ortholog, Cef1p, are essential Myb-related proteins implicated in pre-mRNA splicing and contained within large multiprotein complexes. Here we describe the tandem affinity purification (TAP) of Cdc5p-and Cef1p-associated complexes. Using transmission electron microscopy, we show that the purified Cdc5p complex is a discrete structure. The components of the S. pombe Cdc5p/S. cerevisiae Cef1p complexes (termed Cwfs or Cwcs, respectively) were identified using direct analysis of large protein complex (DALPC) mass spectrometry (A. J. Link et al., Nat. Biotechnol. 17:676-682, 1999). At least 26 proteins were detected in the Cdc5p/Cef1p complexes. Comparison of the polypeptides identified by S. pombe Cdc5p purification with those identified by S. cerevisiae Cef1p purification indicates that these two yeast complexes are nearly identical in composition. The majority of S. pombe Cwf proteins and S. cerevisiae Cwc proteins are known pre-mRNA splicing factors including core Sm and U2 and U5 snRNP components. In addition, the complex contains the U2, U5, and U6 snRNAs. Previously uncharacterized proteins were also identified, and we provide evidence that several of these novel factors are involved in pre-mRNA splicing. Our data represent the first comprehensive analysis of CDC5-associated proteins in yeasts, describe a discrete highly conserved complex containing novel pre-mRNA splicing factors, and demonstrate the power of DALPC for identification of components in multiprotein complexes.The spliceosome is a dynamic multiprotein/RNA complex that catalyzes the excision of introns from pre-mRNA. premRNA splicing requires five snRNA molecules (U1, U2, U4, U5, and U6) that are closely associated with conserved protein components. These snRNA/protein combinations are referred to as small nuclear ribonucleoprotein particles (snRNPs). snRNPs interact with each other and mRNAs in a sequential order during the splicing reaction. The current model for snRNP assembly into an active spliceosome was developed from in vitro splicing experiments (reviewed in references 36, 46, and 59). In addition to the five core snRNP complexes, pre-mRNA splicing requires many other protein factors. Learning the identity of these proteins and defining their roles will further our understanding of how cells regulate the important cellular process of pre-mRNA splicing.The Schizosaccharomyces pombe cdc5 ϩ gene was identified in the first screen for fission yeast mutants defective for cell cycle progression (50). At the restrictive temperature, cells harboring the temperature-sensitive cdc5-120 mutation become arrested in the G 2 phase of the cycle. The cloning and initial characterization of cdc5 ϩ showed that its function is essential for viability and that its predicted protein product shares significant homology with the DNA binding domain of the vertebrate proto-oncoprotein c-Myb (52). Sequence similarity to a family of known DNA binding proteins led us to initially suggest that S. pombe...

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Connections between Pre-mRNA Processing and Regulation of the Eukaryotic Cell Cycle

Cited by 10 publications

References 107 publications

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

Thr¹⁹⁹ phosphorylation targets nucleophosmin to nuclear speckles and represses pre‐mRNA processing

Proteomics Analysis Reveals Stable Multiprotein Complexes in Both Fission and Budding Yeasts Containing Myb-Related Cdc5p/Cef1p, Novel Pre-mRNA Splicing Factors, and snRNAs

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Connections between Pre-mRNA Processing and Regulation of the Eukaryotic Cell Cycle

Cited by 10 publications

References 107 publications

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

Thr199 phosphorylation targets nucleophosmin to nuclear speckles and represses pre‐mRNA processing

Proteomics Analysis Reveals Stable Multiprotein Complexes in Both Fission and Budding Yeasts Containing Myb-Related Cdc5p/Cef1p, Novel Pre-mRNA Splicing Factors, and snRNAs

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Thr¹⁹⁹ phosphorylation targets nucleophosmin to nuclear speckles and represses pre‐mRNA processing