5′-Capping enzymes are targeted to pre-mRNA by binding to the phosphorylated carboxy-terminal domain of RNA polymerase II

McCracken, Susan; Fong, Nova; Rosonina, Emanuel; Yankulov, Krassimir; Siderovski, David P.; Hessel, Andrew; Foster, S. P.; Program, Amgen Est; Shuman, Stewart; Bentley, David L.

doi:10.1101/gad.11.24.3306

Cited by 486 publications

(495 citation statements)

References 55 publications

(84 reference statements)

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“…The phosphorylated CTD binds the capping enzymes (Cho et al 1997;McCracken et al 1997a). CTD overexpression and CTD peptide addition to extracts (Yuryev et al 1996) influence splicing in HeLa cells, which, together with observations that the hyperphosphorylated polymerase II binds snRNPs and SR proteins (Chabot et al 1995;Mortarillo et al 1996;Vincent et al 1996;Kim et al 1997), indicates that spliceosomal components are also associated with RNA polymerase II.…”

mentioning

confidence: 85%

Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3′-end formation and excision of the 3′-terminal intron

Baurén

Беликов²,

Wieslander³

1998

Genes Dev.

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We have analyzed transcription termination, 3 -end formation, and excision of the 3 -terminal intron in vivo in the Balbiani ring 1 (BR1) gene and its pre-mRNA. We show that full-length RNA transcripts are evenly spaced on the gene from a position 300 bp upstream to a region 500-700 bp downstream of the polyadenylation sequence. Very few full-length transcripts and no short, cleaved, nascent transcripts could be observed downstream of this region. Pre-mRNA with 10-20 adenylate residues accumulates at the active gene and then rapidly leaves from the gene locus. Only polyadenylated pre-mRNAs could be detected in the nucleoplasm. Our results are consistent with the hypothesis that transcription termination occurs in a narrow region for the majority of transcripts, simultaneous with 3 -end formation. Excision of the 3 -terminal intron occurs before 3 -end formation in about 5% of the nascent transcripts. When transcription terminates, 3 cleavage takes place and 10-20 adenylate residues are added, the 3 -terminal intron is excised from additionally about 75% of the pre-mRNA at the gene locus. Our data support a close temporal and spatial coupling of transcription termination and the cleavage and initial polyadenylation of 3 -end formation. Excision of the 3 -terminal intron is highly stimulated as the cleavage/polyadenylation complex assembles and 3 -end formation is initiated.

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mentioning

confidence: 85%

Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3′-end formation and excision of the 3′-terminal intron

Baurén

Беликов²,

Wieslander³

1998

Genes Dev.

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“…4 The number of these repeats varies among species with mammals having 52 and C. elegans having 35. 5 The CTD mediates coupling of transcription with pre-mRNA processing [6][7][8][9][10] by acting as a "landing pad" for recruitment of RNA processing factors to the transcription site, 11 thus facilitating co-transcriptional pre-mRNA processing (For a review see refs. [12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Localization of RNAPII and 3′ end formation factor CstF subunits onC. elegansgenes and operons

2016

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Transcription termination is mechanistically coupled to pre-mRNA 3 0 end formation to prevent transcription much beyond the gene 3 0 end. C. elegans, however, engages in polycistronic transcription of operons in which 3 0 end formation between genes is not accompanied by termination. We have performed RNA polymerase II (RNAPII) and CstF ChIP-seq experiments to investigate at a genome-wide level how RNAPII can transcribe through multiple poly-A signals without causing termination. Our data shows that transcription proceeds in some ways as if operons were composed of multiple adjacent single genes. Total RNAPII shows a small peak at the promoter of the gene cluster and a much larger peak at 3 0 ends. These 3 0 peaks coincide with maximal phosphorylation of Ser2 within the C-terminal domain (CTD) of RNAPII and maximal localization of the 3 0 end formation factor CstF. This pattern occurs at all 3 0 ends including those at internal sites in operons where termination does not occur. Thus the normal mechanism of 3 0 end formation does not always result in transcription termination. Furthermore, reduction of CstF50 by RNAi did not substantially alter the pattern of CstF64, total RNAPII, or Ser2 phosphorylation at either internal or terminal 3 0 ends. However, CstF50 RNAi did result in a subtle reduction of CstF64 binding upstream of the site of 3 0 cleavage, suggesting that the CstF50/CTD interaction may facilitate bringing the 3 0 end machinery to the transcription complex.

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“…Srb7 also associates with Tup1, and disruption of the Tup1-Srb7 interaction causes derepression of a target gene (19). The mRNA capping enzyme in S. cerevisiae is composed of Cet1 and Ceg1 (mRNA guanylyltransferase) and is recruited to the phosphorylated C-terminal domain of the largest subunit of RNA polymerase II (54,55). Therefore, at least three proteins associated with RNA polymerase II appear to be targeted by Tup1.…”

Section: Discussionmentioning

confidence: 99%

“…mRNA capping is considered a co-transcriptional event rather than a post-transcriptional event because it occurs when the transcript is only 25-30 nucleotides long (56,57), coincident with the time of C-terminal domain phosphorylation and recruitment of the capping enzyme (54,55). Interestingly the capping enzymes are also bound to and up-regulated by the elongation factor Spt5 and HIV (human immunodeficiency virus) Tat protein (58,59).…”

Section: Discussionmentioning

confidence: 99%

Physical and Functional Interaction of the Yeast Corepressor Tup1 with mRNA 5′-Triphosphatase

Mukai

Davie

Dent

2003

Journal of Biological Chemistry

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The Tup1-Ssn6 complex is an important corepressor in Saccharomyces cerevisiae that inhibits transcription through interactions with the basal transcription machinery and by remodeling chromatin. In a two-hybrid screen for factors that interact with the Schizosaccharomyces pombe Tup1 ortholog, Tup11, we isolated the pct1 ؉ cDNA. The pct1 ؉ gene encodes an mRNA 5 -triphosphatase, which catalyzes the first step of mRNA capping reactions. Pct1 did not interact with the S. pombe Ssn6 ortholog. In vitro glutathione S-transferase pull-down experiments revealed that Pct1 binds to the WD repeat regions of Tup11 and the functionally redundant Tup12 protein. Similarly, the S. cerevisiae Tup1 protein associates with the mRNA 5 -triphosphatase encoded by the CET1 gene. The highly conserved C-terminal domain of Cet1 interacts with Tup1 in vitro, and Tup1-Ssn6 complexes co-purify with the Cet1 protein, indicating that in vivo interactions also occur between these proteins. Over-expression of CET1 compromised repression of an MFA2-lacZ reporter gene that is subject to Tup1-Ssn6 repression. These genetic and biochemical interactions between Tup1-Ssn6 and Cet1 indicate that the capping enzyme associated with RNA polymerase II is a target of the corepressor complex.

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5′-Capping enzymes are targeted to pre-mRNA by binding to the phosphorylated carboxy-terminal domain of RNA polymerase II

Cited by 486 publications

References 55 publications

Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3′-end formation and excision of the 3′-terminal intron

Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3′-end formation and excision of the 3′-terminal intron

Localization of RNAPII and 3′ end formation factor CstF subunits onC. elegansgenes and operons

Physical and Functional Interaction of the Yeast Corepressor Tup1 with mRNA 5′-Triphosphatase

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