Article

Evolutionarily Conserved Interaction between CstF-64 and PC4 Links Transcription, Polyadenylation, and Termination

Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
Molecular Cell (Impact Factor: 14.46). 06/2001; 7(5):1013-23. DOI: 10.1016/S1097-2765(01)00236-2
Source: PubMed

ABSTRACT Tight connections exist between transcription and subsequent processing of mRNA precursors, and interactions between the transcription and polyadenylation machineries seem especially extensive. Using a yeast two-hybrid screen to identify factors that interact with the polyadenylation factor CstF-64, we uncovered an interaction with the transcriptional coactivator PC4. Both human proteins have yeast homologs, Rna15p and Sub1p, respectively, and we show that these two proteins also interact. Given evidence that certain polyadenylation factors, including Rna15p, are necessary for termination in yeast, we show that deletion or overexpression of SUB1 suppresses or enhances, respectively, both growth and termination defects detected in an rna15 mutant strain. Our findings provide an additional, unexpected connection between transcription and polyadenylation and suggest that PC4/Sub1p, via its interaction with CstF-64/Rna15p, possesses an evolutionarily conserved antitermination activity.

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    • "Moreover, according to our protein–protein interaction analysis, TvCstF77, and TvCstF64 interact with TvCPSF160 and TvPC4, respectively ; for this reason these proteins were included in the model. It important to mention that these interactions were previously reported (Calvo and Manley 2001; Zhao et al. 1999). Since Clp1 interacts with PAP and CFI25 (de Vries et al. 2000), so TvClp1 was included in the model. "
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    Genes & genomics 02/2015; 37(4). DOI:10.1007/s13258-015-0268-3 · 0.57 Impact Factor
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    • "Twenty-five milliliters of culture at an OD 600 of 0.3–4 growing at 30ºC and induced with 2% galactose was permeabilized and nascent RNA labeled with [α- 32 P]UTP for 5 min at 30ºC as described (Birse et al., 1998; Calvo and Manley, 2001). After partial hydrolysis, RNA was hybridized directly to filter immobilized YLR454 probes. "
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    • "The allosteric model proposes that the presence of a polyadenylation sequence on the RNA triggers a change in the factors associated with the polymerase [26]. In this model, binding of the cleavage and polyadenylation specificity factors (CPSFs) and the cleavage stimulation factors (CstFs) to the AAUAAA polyadenylation signal on the nascent pre-mRNA favors transcription termination by displacing elongation factors and consequently rendering Pol II less processive [24], [27], [28], [29]. In the torpedo model, the cleavage event at the polyadenylation site generates a new 5′ end [30]. "
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