PXD003619

PXD003619 is an original dataset announced via ProteomeXchange.

Title	The Core RNAi Machinery is Conserved Between the Cytoplasm and Nuclei of Human Cells
Description	Argonaute 2 (AGO2), the catalytic engine of RNAi, is typically associated with inhibition of translation in the cytoplasm. Contrary to dogma, AGO2 has also been implicated in nuclear processes including transcription and splicing. There has been little insight into AGO2's nuclear interactions or how they might differ relative to cytoplasm. Here we investigate the interactions of cytoplasmic and nuclear AGO2 using semi-quantitative mass spectrometry. Mass spectrometry often reveals long lists of candidate proteins, complicating efforts to rigorously discriminate true interacting partners from artifacts. We prioritized candidates using orthogonal analytical strategies that compare replicate mass spectra of proteins associated with FLAG-tagged and endogenous AGO2. Interactions with TRNC6A, TRNC6B, TNRC6C, and AGO3 are conserved between nuclei and cytoplasm. TAR binding protein interacted stably with cytoplasmic AGO2 but not nuclear AGO2, consistent with strand loading in the cytoplasm. Our data suggest that the core RNAi machinery is conserved between the nucleus and cytoplasm but that accessory proteins differ. Orthogonal analysis of mass spectra is a powerful approach to streamlining identification of protein partners.
HostingRepository	MassIVE
AnnounceDate	2016-05-09
AnnouncementXML	Submission_2016-05-09_09:51:27.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andrew Lemoff
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	instrument model: Orbitrap Elite; instrument model: QExactive Plus; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2016-02-11 12:59:37	ID requested
⏵ 1	2016-05-09 09:51:27	announced

Dataset with its publication pending

submitter keyword: Argonaute2, RNAi, Proteomics, Nucleus, Mass Spectrometry

David Corey
lab head
Andrew Lemoff
contact affiliation	UT-Southwestern Medical Center
contact email	andrew.lemoff@utsw.edu
dataset submitter

MassIVE dataset URI

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