Expanding the substantial interactome of NEMO using protein microarrays.

Hdl Handle:
http://hdl.handle.net/10147/93877
Title:
Expanding the substantial interactome of NEMO using protein microarrays.
Authors:
Fenner, Beau J; Scannell, Michael; Prehn, Jochen H M
Affiliation:
Centre for Human Proteomics and Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland.
Citation:
Expanding the substantial interactome of NEMO using protein microarrays. 2010, 5 (1):e8799 PLoS ONE
Journal:
PloS one
Issue Date:
2010
URI:
http://hdl.handle.net/10147/93877
DOI:
10.1371/journal.pone.0008799
PubMed ID:
20098747
Abstract:
Signal transduction by the NF-kappaB pathway is a key regulator of a host of cellular responses to extracellular and intracellular messages. The NEMO adaptor protein lies at the top of this pathway and serves as a molecular conduit, connecting signals transmitted from upstream sensors to the downstream NF-kappaB transcription factor and subsequent gene activation. The position of NEMO within this pathway makes it an attractive target from which to search for new proteins that link NF-kappaB signaling to additional pathways and upstream effectors. In this work, we have used protein microarrays to identify novel NEMO interactors. A total of 112 protein interactors were identified, with the most statistically significant hit being the canonical NEMO interactor IKKbeta, with IKKalpha also being identified. Of the novel interactors, more than 30% were kinases, while at least 25% were involved in signal transduction. Binding of NEMO to several interactors, including CALB1, CDK2, SAG, SENP2 and SYT1, was confirmed using GST pulldown assays and coimmunoprecipitation, validating the initial screening approach. Overexpression of CALB1, CDK2 and SAG was found to stimulate transcriptional activation by NF-kappaB, while SYT1 overexpression repressed TNFalpha-dependent NF-kappaB transcriptional activation in human embryonic kidney cells. Corresponding with this finding, RNA silencing of CDK2, SAG and SENP2 reduced NF-kappaB transcriptional activation, supporting a positive role for these proteins in the NF-kappaB pathway. The identification of a host of new NEMO interactors opens up new research opportunities to improve understanding of this essential cell signaling pathway.
Language:
en
ISSN:
1932-6203

Full metadata record

DC FieldValue Language
dc.contributor.authorFenner, Beau Jen
dc.contributor.authorScannell, Michaelen
dc.contributor.authorPrehn, Jochen H Men
dc.date.accessioned2010-03-08T14:44:53Z-
dc.date.available2010-03-08T14:44:53Z-
dc.date.issued2010-
dc.identifier.citationExpanding the substantial interactome of NEMO using protein microarrays. 2010, 5 (1):e8799 PLoS ONEen
dc.identifier.issn1932-6203-
dc.identifier.pmid20098747-
dc.identifier.doi10.1371/journal.pone.0008799-
dc.identifier.urihttp://hdl.handle.net/10147/93877-
dc.description.abstractSignal transduction by the NF-kappaB pathway is a key regulator of a host of cellular responses to extracellular and intracellular messages. The NEMO adaptor protein lies at the top of this pathway and serves as a molecular conduit, connecting signals transmitted from upstream sensors to the downstream NF-kappaB transcription factor and subsequent gene activation. The position of NEMO within this pathway makes it an attractive target from which to search for new proteins that link NF-kappaB signaling to additional pathways and upstream effectors. In this work, we have used protein microarrays to identify novel NEMO interactors. A total of 112 protein interactors were identified, with the most statistically significant hit being the canonical NEMO interactor IKKbeta, with IKKalpha also being identified. Of the novel interactors, more than 30% were kinases, while at least 25% were involved in signal transduction. Binding of NEMO to several interactors, including CALB1, CDK2, SAG, SENP2 and SYT1, was confirmed using GST pulldown assays and coimmunoprecipitation, validating the initial screening approach. Overexpression of CALB1, CDK2 and SAG was found to stimulate transcriptional activation by NF-kappaB, while SYT1 overexpression repressed TNFalpha-dependent NF-kappaB transcriptional activation in human embryonic kidney cells. Corresponding with this finding, RNA silencing of CDK2, SAG and SENP2 reduced NF-kappaB transcriptional activation, supporting a positive role for these proteins in the NF-kappaB pathway. The identification of a host of new NEMO interactors opens up new research opportunities to improve understanding of this essential cell signaling pathway.-
dc.language.isoenen
dc.titleExpanding the substantial interactome of NEMO using protein microarrays.en
dc.contributor.departmentCentre for Human Proteomics and Department of Physiology and Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland.en
dc.identifier.journalPloS oneen

Related articles on PubMed

All Items in Lenus, The Irish Health Repository are protected by copyright, with all rights reserved, unless otherwise indicated.