A Large Scale shRNA Barcode Screen Identifies the Circadian Clock Component ARNTL as Putative Regulator of the p53 Tumor Suppressor Pathway

The p53 tumor suppressor gene is mutated in about half of human cancers, but the p53 pathway is thought to be functionally inactivated in the vast majority of cancer. Understanding how tumor cells can become insensitive to p53 activation is therefore of major importance. Using an RNAi-based genetic...

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Veröffentlicht in:	PloS one Jg. 4; H. 3; S. e4798
Hauptverfasser:	Mullenders, Jasper, Fabius, Armida W. M., Madiredjo, Mandy, Bernards, René, Beijersbergen, Roderick L.
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	United States Public Library of Science 11.03.2009 Public Library of Science (PLoS)
Schlagworte:	Activation Antigens ARNTL Transcription Factors Bar codes Basic Helix-Loop-Helix Transcription Factors - genetics Basic Helix-Loop-Helix Transcription Factors - physiology BMAL1 protein Cancer Cell Biology/Cell Signaling Cell Biology/Cellular Death and Stress Responses Cell Biology/Gene Expression Cell cycle Cell Division Cell Line Circadian rhythm Circadian Rhythm - genetics Circadian rhythms Cyclin-dependent kinase inhibitor p21 Cyclin-Dependent Kinase Inhibitor p21 - biosynthesis Cyclin-Dependent Kinase Inhibitor p21 - genetics Cytokines Deoxyribonucleic acid DNA DNA damage DNA methylation DNA-Binding Proteins - genetics DNA-Binding Proteins - physiology Gene Library Genes Genes, p53 Genetic screening Genetic testing Genetic Vectors - genetics Genetics Genomics Humans Inverted Repeat Sequences Medical screening Modulators Molecular Biology Oligonucleotide Probes Oncology p53 Protein Population Proteins Regulators RNA Interference RNA, Small Interfering - genetics RNA, Small Interfering - isolation & purification RNA-mediated interference Transcription Factors - genetics Transcription Factors - physiology Tumor cells Tumor necrosis factor-TNF Tumor proteins Tumor suppressor genes Tumor Suppressor Protein p53 - physiology Ubiquitin-Protein Ligases
ISSN:	1932-6203, 1932-6203
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Zusammenfassung:	The p53 tumor suppressor gene is mutated in about half of human cancers, but the p53 pathway is thought to be functionally inactivated in the vast majority of cancer. Understanding how tumor cells can become insensitive to p53 activation is therefore of major importance. Using an RNAi-based genetic screen, we have identified three novel genes that regulate p53 function. We have screened the NKI shRNA library targeting 8,000 human genes to identify modulators of p53 function. Using the shRNA barcode technique we were able to quickly identify active shRNA vectors from a complex mixture. Validation of the screening results indicates that the shRNA barcode technique can reliable identify active shRNA vectors from a complex pool. Using this approach we have identified three genes, ARNTL, RBCK1 and TNIP1, previously unknown to regulate p53 function. Importantly, ARNTL (BMAL1) is an established component of the circadian regulatory network. The latter finding adds to recent observations that link circadian rhythm to the cell cycle and cancer. We show that cells having suppressed ARNTL are unable to arrest upon p53 activation associated with an inability to activate the p53 target gene p21(CIP1). We identified three new regulators of the p53 pathway through a functional genetic screen. The identification of the circadian core component ARNTL strengthens the link between circadian rhythm and cancer.
Bibliographie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Conceived and designed the experiments: JM AWF RB RB. Performed the experiments: JM AWF MM. Analyzed the data: JM AWF RB. Wrote the paper: JM AWF RB RB. Performed the microarray hybridizations: MM. Supervised the research and corrected the manuscript: RB RLB.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0004798