Polyadenylate Polymerase (PAP) and 3' End pre-mRNA Processing: Function, Assays, and Association with Disease

Scorilas, Andreas

doi:10.1080/10408360290795510

Cited by 29 publications

(22 citation statements)

References 176 publications

(221 reference statements)

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“…SUMO has a substantial impact on PAP function, and our data indicate that alterations in PAP sumoylation have the potential to profoundly affect gene expression. For example, misregulation of PAP activity has been associated with disease (for review, see Scorilas 2002). PAP activity is known to reflect the proliferative capacity of cells, as several neoplastic and leukaemic cells show increased PAP activity with or without increased PAP expression (Papamichail et al 1983).…”

Section: Discussionmentioning

confidence: 99%

Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function

Vethantham¹,

Rao²,

Manley³

2008

Genes Dev.

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The addition of the poly(A) tail to the ends of eukaryotic mRNAs is catalyzed by poly(A) polymerase (PAP).PAP activity is known to be highly regulated, for example, by alternative splicing and phosphorylation. In this study we show that the small ubiquitin-like modifier (SUMO) plays multiple roles in regulating PAP function. Our discovery of SUMO-conjugated PAP began with the observation of a striking pattern of abundant higher-molecular-weight forms of PAP in certain mouse tissues and cell lines. PAP constitutes an unusual SUMO substrate in that, despite the absence of any consensus sumoylation sites, PAP interacts very strongly with the SUMO E2 enzyme ubc9 and can be extensively sumoylated both in vitro and in vivo. Six sites of sumoylation in PAP were identified, with two overlapping one of two nuclear localization signals (NLS). Strikingly, mutation of the two lysines at the NLS to arginines, or coexpression of a SUMO protease with wild-type PAP, caused PAP to be localized to the cytoplasm, demonstrating that sumoylation is required to facilitate PAP nuclear localization. Sumoylation also contributes to PAP stability, as down-regulation of sumoylation led to decreases in PAP levels. Finally, the activity of purified PAP was shown to be inhibited by in vitro sumoylation. Our study thus shows that SUMO regulates PAP in numerous distinct ways and is integral to normal PAP function.[Keywords: PAP; sumoylation; polyadenylation; ubc9; localization] Supplemental material is available at http://www.genesdev.org.

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Section: Discussionmentioning

confidence: 99%

Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function

Vethantham¹,

Rao²,

Manley³

2008

Genes Dev.

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“…The assembly of the cleavage/polyadenylation machinery requires specific signal sequences in the mRNA precursor as well as interactions of a large number of protein factors (reviewed by Mandel et al, 2008). It has been shown that the regulation of mRNA 3 0 end formation can have significant roles in cancer (Kleiman and Manley, 2001;Topalian et al, 2001;Scorilas, 2002;Rozenblatt-Rosen et al, 2009). Most importantly, alternative mRNA cleavage and polyadenylation changes the length of the 3 0 -untranslated region and regulates gene expression of different mRNAs in cancer cells (Mayr and Bartel 2009;Singh et al, 2009) and during cell differentiation (Sandberg et al, 2008;Zlotorynski and Agami, 2008;Ji et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

p53 inhibits mRNA 3′ processing through its interaction with the CstF/BARD1 complex

et al. 2011

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The mechanisms involved in the p53-dependent control of gene expression following DNA damage have not been completely elucidated. Here, we show that the p53 C terminus associates with factors that are required for the ultraviolet (UV)-induced inhibition of the mRNA 3 0 cleavage step of the polyadenylation reaction, such as the tumor suppressor BARD1 and the 3 0 processing factor cleavage-stimulation factor 1 (CstF1). We found that p53 can coexist in complexes with CstF and BARD1 in extracts of UV-treated cells, suggesting a role for p53 in mRNA 3 0 cleavage following DNA damage. Consistent with this, we found that p53 inhibits 3 0 cleavage in vitro and that there is a reverse correlation between the levels of p53 expression and the levels of mRNA 3 0 cleavage under different cellular conditions. Supporting these results, a tumor-associated mutation in p53 not only decreases the interaction with BARD1 and CstF, but also decreases the UV-induced inhibition of 3 0 processing, all of which is restored by wild-type-p53 expression. We also found that p53 expression levels affect the polyadenylation levels of housekeeping genes, but not of p21 and c-fos genes, which are involved in the DNA damage response (DDR). Here, we identify a novel 3 0 RNA processing inhibitory function of p53, adding a new level of complexity to the DDR by linking RNA processing to the p53 network.

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“…19,20 The cloned sequences of bovine and human eNOS mRNAs were reported to have 3Ј poly(A) tails Ϸ12 to 20 nucleotides in length. 21,22 Although these studies did not specifically examine poly(A) tail length, the eNOS sequences were derived from RNA libraries of cultured cells not exposed to shear, suggesting that eNOS mRNA has a short 3Ј poly(A) tail under baseline conditions.…”

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confidence: 99%

Laminar Shear Stress and 3′ Polyadenylation of eNOS mRNA

Weber

Hagedorn

Harrison

et al. 2005

Circulation Research

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Abstract-The 3Ј poly(A) tail is important in messenger RNA stability and translational efficiency. In somatic tissues, 3Ј polyadenylation of mRNAs has been thought to largely be a constitutively active process. We have reported that laminar shear stress causes a brief increase in endothelial nitric oxide synthase (eNOS) transcription, followed by a prolonged increase in eNOS mRNA stability. We sought to determine whether shear stress and other stimuli affected eNOS 3Ј polyadenylation in endothelial cells. Under basal (static) conditions, eNOS mRNA possessed short 3Ј poly(A) tails of Ͻ25 nt. In contrast, laminar shear stress increased expression of eNOS transcripts with long poly(A) tails. ENOS transcripts with longer poly(A) tails had prolonged half-lives (6 hours in static cells versus 18 hours in sheared cells). Polysome analysis revealed that eNOS mRNA from sheared cells was shifted into more translationally active polysome fractions compared with eNOS mRNA from static cells. Shear-induced lengthening of the eNOS 3Ј poly(A) tail was the result of increased nuclear polyadenylation. Furthermore, hydrogen peroxide and HMG Co-A reductase inhibitors, other stimuli known to modulate eNOS expression posttranscriptionally, also induced eNOS 3Ј poly(A) tail lengthening. These results support the concept that shear stress modulates eNOS mRNA stability and translation via increased 3Ј polyadenylation. We suggest that mRNA 3Ј polyadenylation is a posttranscriptional mechanism used by endothelial cells to regulate gene expression. Key Words: endothelial nitric oxide synthase Ⅲ mRNA stability Ⅲ polyadenylation Ⅲ posttranscriptional regulation Ⅲ shear stress L aminar shear stress is a potent stimulus for the production of vascular nitric oxide (NO • ) because of its ability to increase both the activity and expression of the endothelial nitric oxide synthase (eNOS). 1-3 Endothelium-derived NO• is crucial for maintenance of vascular homeostasis through its vasodilator activity; 4 its ability to inhibit smooth muscle growth, 5 platelet aggregation, 6 and leukocyte adhesion, 7 and its role in inhibiting lipid oxidation and regulating apoptosis in the vessel wall. 8 Given these properties of NO• , the shear stress-induced increase in eNOS expression may be important in preventing atherosclerosis. Indeed, areas of the vasculature exposed to high shear stress appear to be protected from the development of atherosclerosis, and areas exposed to low shear stress are prone to atherosclerotic lesion formation. 9 Previously we have demonstrated that shear stress leads to increased eNOS mRNA expression via 2 separate mechanisms: a transient increase in eNOS transcription, and stabilization of eNOS mRNA. 10 Posttranscriptional regulation of eNOS expression via modulation of eNOS mRNA stability is now recognized as an important response of endothelial cells to numerous biophysical and biochemical stimuli. 11-17 Although we have recently described a mechanism for the posttranscriptional regulation of eNOS expression during cell growth, 18 ...

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Polyadenylate Polymerase (PAP) and 3' End pre-mRNA Processing: Function, Assays, and Association with Disease

Cited by 29 publications

References 176 publications

Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function

Sumoylation regulates multiple aspects of mammalian poly(A) polymerase function

p53 inhibits mRNA 3′ processing through its interaction with the CstF/BARD1 complex

Laminar Shear Stress and 3′ Polyadenylation of eNOS mRNA

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