Cleavage Factor II of Saccharomyces cerevisiaeContains Homologues to Subunits of the Mammalian Cleavage/ Polyadenylation Specificity Factor and Exhibits Sequence-specific, ATP-dependent Interaction with Precursor RNA

Zhao, Jing; Kessler, Marco M.; Moore, Claire

doi:10.1074/jbc.272.16.10831

Cited by 57 publications

(86 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We also checked for the presence of the Clp1p subunit of CF IA in the eluates, but the low affinity of this antibody precluded the detection of this protein in the pellet fraction. However, given the tight association of the subunits of CF IA (17)(18)(19)(20)(21)(22)(23), Clp1p is likely to be recruited to the CTD as part of the CF IA complex. The interaction observed was resistant to treatment with RNase, arguing against the possibility of a RNA bridge (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…To date the interaction of cleavage͞polyadenylation factors with the CTD remains largely uncharacterized. In yeast 3Ј-end processing of pre-mRNA requires cleavage factor IA (CF IA), cleavage factor IB (CF IB), cleavage factor II (CF II), polyadenylation factor I (PFI), poly(A) polymerase, and the poly(A)-binding protein 1 (17)(18)(19)(20)(21)(22). CF IA, which is involved in both cleavage and polyadenylation, comprises four polypeptides: Rna15p, Rna14p, Pcf11p, and Clp1p.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Cleavage/polyadenylation factor IA associates with the carboxyl-terminal domain of RNA polymerase II in Saccharomycescerevisiae

Barillà¹

2001

Proceedings of the National Academy of Sciences

View full text Add to dashboard Cite

The carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II plays an important role in transcription and processing of the nascent transcript by interacting with both transcription and RNA processing factors. We show here that the cleavage͞ polyadenylation factor IA of Saccharomyces cerevisiae directly contacts CTD. First by affinity chromatography experiments with yeast extracts we demonstrate that the Rna15p, Rna14p, and Pcf11p subunits of this complex are associated with phosphorylated CTD. This interaction is confirmed for Rna15p by yeast two-hybrid analysis. Second, Pcf11p, but not Rna15p, is shown to directly contact phosphorylated CTD based on in vitro binding studies with recombinant proteins. These findings establish a direct interaction of cleavage͞polyadenylation factor IA with the CTD. Furthermore, a quantitative analysis of transcription run-on performed on temperature-sensitive mutant strains reveals that the lack of either functional Rna14p or Pcf11p affects transcription termination more severely than the absence of a functional Rna15p. Moreover, these data reinforce the concept that CTD phosphorylation acts as a regulatory mechanism in the maturation of the primary transcript. In Saccharomyces cerevisiae the largest subunit of RNA polymerase II (pol II) contains a carboxyl-terminal domain (CTD) consisting of 26 tandem repeats of a heptapeptide module with the consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser (1, 2). Two serines per heptad repeat may undergo reversible phosphorylation during each transcription cycle (3). The phosphorylation status of the CTD is correlated with different stages of the transcription cycle. Thus hypophosphorylated polymerase (IIA) is competent for initiation, whereas hyperphosphorylated polymerase (II0) is associated with transcription elongation. Increasing evidence has been provided in the last few years showing that the CTD acts as a direct physical link between transcription and nascent RNA processing: in mammals, the cleavage-polyadenylation specificity factor (CPSF) and the cleavage stimulation factor (CstF), as well as splicing factors and 5Ј capping enzyme, all bind to the CTD (4-9). Furthermore, CPSF and CstF copurify with pol II (4). The concept of a factor recruiting͞docking platform has emerged as one likely function of this peculiar polypeptide. Evidence for a more direct role of the CTD in polyadenylation has also been indicated (10).In S. cerevisiae, the proteins so far known to bind to the CTD are the Mediator complex (11, 12), the Elongator complex (13), proteins involved in 5Ј end capping of nascent RNA (8,14), and the Ess1 peptidylprolyl isomerase (15, 16). To date the interaction of cleavage͞polyadenylation factors with the CTD remains largely uncharacterized. In yeast 3Ј-end processing of pre-mRNA requires cleavage factor IA (CF IA), cleavage factor IB (CF IB), cleavage factor II (CF II), polyadenylation factor I (PFI), poly(A) polymerase, and the poly(A)-binding protein 1 (17-22). CF IA, which is involved in both cleavage and po...

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Cleavage/polyadenylation factor IA associates with the carboxyl-terminal domain of RNA polymerase II in Saccharomycescerevisiae

Barillà¹

2001

Proceedings of the National Academy of Sciences

View full text Add to dashboard Cite

show abstract

“…Yeast two-hybrid studies showed that the last 245 residues of CPSF-73 can interact with CPSF-100 [117]. Besides protein-protein interactions, Cft2p/Ydh1p can bind the pre-mRNA near the cleavage site [72,122].…”

Section: Cpsf-100 (Cft2p/ydh1p)mentioning

confidence: 99%

“…Interestingly, Yth1p does not appear to co-purify with the other CPSF subunits (Cft1p/Yhh1p, Cft2p/Ydh1p, Brr5p/ Ysh1p, Fip1p), even though they are in the larger CPF complex (Fig. 1B) [115,122]. CPSF-30 has a second isoform in humans (locus XP_945726), sharing 54% sequence identity.…”

Section: Cpsf-30 (Yth1p)mentioning

confidence: 99%

Protein factors in pre-mRNA 3′-end processing

Mandel

Bai

Tong

2007

Cell. Mol. Life Sci.

364

482

View full text Add to dashboard Cite

Most eukaryotic mRNA precursors (pre-mRNAs) must undergo extensive processing, including cleavage and polyadenylation at the 3′-end. Processing at the 3′-end is controlled by sequence elements in the pre-mRNA (cis elements) as well as protein factors. Despite the seeming biochemical simplicity of the processing reactions, more than 14 proteins have been identified for the mammalian complex, and more than 20 proteins have been identified for the yeast complex. The 3′-end processing machinery also has important roles in transcription and splicing. The mammalian machinery contains several sub-complexes, including cleavage and polyadenylation specificity factor (CPSF), cleavage stimulation factor (CstF), cleavage factor I (CF I m ), and cleavage factor II (CF II m ). Additional protein factors include poly(A) polymerase (PAP), poly(A) binding protein (PABP), symplekin, and the C-terminal domain (CTD) of RNA polymerase II largest subunit. The yeast machinery includes cleavage factor IA (CF IA), cleavage factor IB (CF IB), and cleavage and polyadenylation factor (CPF).

show abstract

“…Further purification of these factors has led toward a determination of the number and identity of their constituent proteins. CF II contains the four peptides Cft1, Cft2, Brr5, and Pta1 (8). PF I contains the CF II subunits and the additional peptides Pfs1, Fip1, Pfs2, and Yth1 (9).…”

mentioning

confidence: 99%

Five subunits are required for reconstitution of the cleavage and polyadenylation activities of Saccharomyces cerevisiae cleavage factor I

Größ

Moore

2001

Proc. Natl. Acad. Sci. U.S.A.

142

View full text Add to dashboard Cite

Cleavage and polyadenylation of mRNA 3 ends in Saccharomyces cerevisiae requires several factors, one of which is cleavage factor I (CF I). Purification of CF I activity from yeast extract has implicated numerous proteins as functioning in both cleavage and͞or polyadenylation. Through reconstitution of active CF I from separately expressed and purified proteins, we show that CF I contains five subunits, Rna14, Rna15, Pcf11, Clp1, and Hrp1. These five are necessary and sufficient for reconstitution of cleavage activity in vitro when mixed with CF II, and for specific polyadenylation when mixed with polyadenylation factor I, purified poly(A) polymerase, and poly(A) binding protein. Analysis of the individual proteinprotein interactions supports an architectural model for CF I in which Pcf11 simultaneously interacts with Rna14, Rna15, and Clp1, whereas Rna14 bridges Rna15 and Hrp1.

show abstract

Cleavage Factor II of Saccharomyces cerevisiaeContains Homologues to Subunits of the Mammalian Cleavage/ Polyadenylation Specificity Factor and Exhibits Sequence-specific, ATP-dependent Interaction with Precursor RNA

Cited by 57 publications

References 47 publications

Cleavage/polyadenylation factor IA associates with the carboxyl-terminal domain of RNA polymerase II in Saccharomycescerevisiae

Cleavage/polyadenylation factor IA associates with the carboxyl-terminal domain of RNA polymerase II in Saccharomycescerevisiae

Protein factors in pre-mRNA 3′-end processing

Five subunits are required for reconstitution of the cleavage and polyadenylation activities of Saccharomyces cerevisiae cleavage factor I

Contact Info

Product

Resources

About