NPAT Expression Is Regulated by E2F and Is Essential for Cell Cycle Progression

Gao, Guang; Bracken, Adrian P.; Burkard, Karina; Pasini, Diego; Classon, Marie; Attwooll, Claire; Sagara, Masashi; Imai, Takashi; Helin, Kristian; Zhao, Jiapeng

doi:10.1128/mcb.23.8.2821-2833.2003

Cited by 57 publications

(73 citation statements)

References 55 publications

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“…Additionally, there was no apparent homogeneity of function among these factors, except that most fitted into the categories related to cell cycle regulation and DNA recombination and repair. Of the five proteins affected by the most effective siRNAs, SETD8 is a SET domain containing lysine methyltransferase (37), CASP8AP2 is a component of the apoptotic machinery (38), SOX15 is a developmental transcription factor (39), TROAP is a cell adhesion molecule reported to mediate blastocyst implantation (40), and NPAT is a cell cycle progression regulator encoded within the ATM gene (41). This heterogeneity of function might indicate that AAV transduction is regulated at multiple levels or that, more likely, the knockdown of these factors converge into common intracellular signaling pathways.…”

Section: Discussionmentioning

confidence: 99%

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

Mano

Ippodrino

Zentilin

et al. 2015

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

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Viral vectors based on the adeno-associated virus (AAV) hold great promise for in vivo gene transfer; several unknowns, however, still limit the vectors' broader and more efficient application. Here, we report the results of a high-throughput, whole-genome siRNA screening aimed at identifying cellular factors regulating AAV transduction. We identified 1,483 genes affecting vector efficiency more than 4-fold and up to 50-fold, either negatively or positively. Most of these factors have not previously been associated to AAV infection. The most effective siRNAs were independent from the virus serotype or analyzed cell type and were equally evident for single-stranded and self-complementary AAV vectors. A common characteristic of the most effective siRNAs was the induction of cellular DNA damage and activation of a cell cycle checkpoint. This information can be exploited for the development of more efficient AAV-based gene delivery procedures. Administration of the most effective siRNAs identified by the screening to the liver significantly improved in vivo AAV transduction efficiency.adeno-associated virus | DNA-damage response | high-throughput screening | self-complementary vectors | RNA interference V iral vectors based on the adeno-associated virus (AAV) have received incremental attention over the past two decades as effective tools for in vivo gene transfer. The vectors' intrinsic structural simplicity, lack of pathogenicity, low immunogenicity, and ability to mediate long-term, episomal expression in nondividing cells in vitro and, most notably, postmitotic organs in vivo are desirable characteristics for several gene therapy applications (reviewed in ref. 1). Indeed, to date, over 90 clinical trials using these vectors have been carried out (www.abedia.com/wiley/index. html); the first commercial gene therapy product (Glybera) is an AAV1 vector for the treatment of familial lipoprotein lipase deficiency (2).Despite these achievements, AAV vectors still display an undeniable number of limitations in terms of restricted cellular permissivity and efficacy of transgene expression. Efficient transduction in vivo is essentially limited to postmitotic cells [in particular, cardiomyocytes, neurons, retinal cells, and skeletal muscle fibers (1)] and requires infection at a relatively high multiplicity of infection; in addition, vector-driven gene expression is achieved only after a relatively long lag period (3). These characteristics are still incompletely understood in molecular terms but certainly reside within the intrinsic biological properties of target cells. This finding is also highlighted by the peculiar biology of wild-type AAV, which, for the completion of its biological cycle, requires cellular coinfection by adenovirus or other viruses that modify the cellular environment of the target cells, rendering them permissive for viral replication (4-6).The efficient internalization of AAV vectors into different cell types is mediated by the binding of virions to ubiquitously expressed cell surface receptors (...

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Section: Discussionmentioning

confidence: 99%

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

Mano

Ippodrino

Zentilin

et al. 2015

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

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“…Nuclear Protein, Ataxia-Telangiectasia Locus (NPAT/ p220) stimulates the transcription of all replicationdependent histone genes during S phase (Gao et al, 2003;Ye et al, 2003) in a phosphorylation-dependent manner (Zhao et al, 1998(Zhao et al, , 2000Ma et al, 2000). NPAT functions, in part, by recruiting coactivator proteins including histone-modifying enzymes (DeRan et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…NPAT functions, in part, by recruiting coactivator proteins including histone-modifying enzymes (DeRan et al, 2008). Moreover, NPAT transcription is regulated in a cell cycle-dependent manner by the E2F family of transcription factors (Gao et al, 2003). However, no role for NPAT in histone mRNA 3 0 end processing has been reported.…”

Section: Introductionmentioning

confidence: 99%

Induced G1 cell-cycle arrest controls replication-dependent histone mRNA 3′ end processing through p21, NPAT and CDK9

Pirngruber

Johnsen

2010

Oncogene

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Proper cell cycle-dependent expression of replicationdependent histones is essential for packaging of DNA into chromatin during replication. We previously showed that cyclin-dependent kinase-9 (CDK9) controls histone H2B monoubiquitination (H2Bub1) to direct the recruitment of specific mRNA 3 0 end processing proteins to replicationdependent histone genes and promote proper pre-mRNA 3 0 end processing. We now show that p53 decreases the expression of the histone-specific transcriptional regulator Nuclear Protein, Ataxia-Telangiectasia Locus (NPAT) by inducing a G1 cell-cycle arrest, thereby affecting E2F-dependent transcription of the NPAT gene. Furthermore, NPAT is essential for histone mRNA 3 0 end processing and recruits CDK9 to replication-dependent histone genes. Reduced NPAT expression following p53 activation or small interfering RNA knockdown decreases CDK9 recruitment and replication-dependent histone gene transcription but increases the polyadenylation of remaining histone mRNAs. Thus, we present evidence that the induction of a G1 cell-cycle arrest (for example, following p53 accumulation) alters histone mRNA 3 0 end processing and uncover the first mechanism of a regulated switch in the mode of pre-mRNA 3 0 end processing during a normal cellular process, which may be altered during tumorigenesis.

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“…NPAT is a newly discovered target gene of E2F-1 (Gao et al, 2003) and we wanted to know if NPAT is also repressed by induced HES-1 expression in T47D cells. By using the T47D dnHES-1 cell line, we found that treatment with atRA prevented E 2 induction of NPAT mRNA, while expression of dnHES-1 reversed the effects of atRA (Figure 4b).…”

Section: Hes-1 Expression Arrests T47d Cells In the G 1 Phase Of The mentioning

confidence: 99%

HES-1 inhibits 17β-estradiol and heregulin-β1-mediated upregulation of E2F-1

et al. 2004

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We have previously shown that expression of the transcription factor HES-1 is required for the growthinhibitory effect of all-trans retinoic acid on MCF-7 cells. In this study, we have used T47D cells with tetracyclinregulated expression of wild-type or a dominant-negative form of HES-1. Expression of HES-1 in T47D cells inhibited G 1 /S-phase transition and activation of Cdk2 elicited by estrogen. Estrogen treatment of T47D cells caused increased expression of E2F-1, and this expression was inhibited by cotreatment with all-trans retinoic acid. We show that the effect is mediated through HES-1, which directly downregulates E2F-1 expression through a CACGAG-site within the E2F-1 promoter. Furthermore, proliferation caused by heregulin-b1 treatment of T47D cells was inhibited by all-trans retinoic acid and this effect was mediated by HES-1. Interestingly, heregulin-b1-mediated upregulation of E2F-1 expression was directly inhibited by HES-1 through the same CACGAG-site as seen with estrogen-stimulated induction. In addition, we found that two important downstream target genes of estrogen and heregulin-b1 that are regulated through E2F-1, cyclin E and NPAT, were both regulated in a similar fashion by all-trans retinoic acid, and these effects were antagonized by dominant-negative HES-1. These findings establish that HES-1 inhibits both estrogen-and heregulin-b1-stimulated growth of breast cancer cells, and further suggest that growth inhibition induced in these cells by all-trans retinoic acid occurs via HES-1-mediated downregulation of E2F-1 expression.

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NPAT Expression Is Regulated by E2F and Is Essential for Cell Cycle Progression

Cited by 57 publications

References 55 publications

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

Induced G1 cell-cycle arrest controls replication-dependent histone mRNA 3′ end processing through p21, NPAT and CDK9

HES-1 inhibits 17β-estradiol and heregulin-β1-mediated upregulation of E2F-1

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