NDLI: Genome-scale RNAi on living-cell microarrays identifies novel regulators of Drosophila melanogaster TORC1-S6K pathway signaling.

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Genome-scale RNAi on living-cell microarrays identifies novel regulators of Drosophila melanogaster TORC1-S6K pathway signaling.

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Genome-scale RNAi on living-cell microarrays identifies novel regulators of Drosophila melanogaster TORC1-S6K pathway signaling.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lindquist, Robert A. Ottina, Kathleen A. Wheeler, Douglas B. Hsu, Peggy P. Thoreen, Carson C. Guertin, David A. Ali, Siraj M. Sengupta, Shomit Shaul, Yoav D. Lamprecht, Michael R. Madden, Katherine L. Papallo, Adam R. Jones, Thouis R. Sabatini, David M. Carpenter, Anne E.
Description	Country affiliation: United States Author Affiliation: Lindquist RA ( Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA.)
Abstract	The evolutionarily conserved target of rapamycin complex 1 (TORC1) controls cell growth in response to nutrient availability and growth factors. TORC1 signaling is hyperactive in cancer, and regulators of TORC1 signaling may represent therapeutic targets for human diseases. To identify novel regulators of TORC1 signaling, we performed a genome-scale RNA interference screen on microarrays of Drosophila melanogaster cells expressing human RPS6, a TORC1 effector whose phosphorylated form we detected by immunofluorescence. Our screen revealed that the TORC1-S6K-RPS6 signaling axis is regulated by many subcellular components, including the Class I vesicle coat (COPI), the spliceosome, the proteasome, the nuclear pore, and the translation initiation machinery. Using additional RNAi reagents, we confirmed 70 novel genes as significant on-target regulators of RPS6 phosphorylation, and we characterized them with extensive secondary assays probing various arms of the TORC1 pathways, identifying functional relationships among those genes. We conclude that cell-based microarrays are a useful platform for genome-scale and secondary screening in Drosophila, revealing regulators that may represent drug targets for cancers and other diseases of deregulated TORC1 signaling.
File Format	HTM / HTML
ISSN	10889051
e-ISSN	15495469
DOI	10.1101/gr.111492.110
Journal	Genome Research
Issue Number	3
Volume Number	21
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2011-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Genetics Discipline Genomics Recombinant Proteins Metabolism Ribosomal Protein S6 Transcription Factors Animals Blotting, Western Cells, Cultured Drosophila Proteins Genetics Drosophila Melanogaster Fluorescent Antibody Technique Gene Regulatory Networks Genome Genomics Microarray Analysis Molecular Targeted Therapy Phosphorylation Rna Interference Signal Transduction Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s.
Content Type	Text
Resource Type	Article
Subject	Genetics Genetics (clinical)

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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Genome-scale RNAi on living-cell microarrays identifies novel regulators of Drosophila melanogaster TORC1–S6K pathway signaling

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