Proteomics

Dataset Information

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Mapping of the nuclear PI(3,4,5)P3 interactome in DLD1 and SW480 cells


ABSTRACT: Phosphatidylinositol (3,4,5)-trisphosphate (PI(3,4,5)P3) is a lipid second messenger produced by class I phosphoinositide 3-kinases (PI 3-kinases) phosphorylation of phosphatidylinositol (4,5)-bisphosphate (PI(4,5)P2). Although typically associated with the plasma membrane, it is now appreciated that PI(3,4,5)P3 generation also occurs in the nucleus to regulate a variety of biological functions. Using a PI(3,4,5)P3-specific GST-GRP1PH domain reporter and immunofluorescence microscopy, we have found that DLD1 and SW480 colorectal cancer (CRC) cells exhibit pronounced nuclear staining of PI(3,4,5)P3. To characterise the nuclear PI(3,4,5)P3 interactome in DLD1 and SW480 cells, we performed affinity pull-down experiments on nuclear extracts using a ω-amino analogue of phosphatidylinositol 3,4,5 -triphosphate (PI(3,4,5)P3 immobilised onto Affi-gel10 beads. Gel fractionation and LC/MS-MS analysis was performed to reduce sample complexity and increase resolution, with samples run on a LTQ Orbitrap Elite mass spectrometer. DLD1 and SW480 cells were both analysed in four independent biological replicates. A protein was considered a member of the nuclear PI(3,4,5)P3 interactome if it was identified in at least two experiments (with at least two peptides, FDR of 1%) in both cell lines, yielding a total of 1237 unique proteins (representing 1103 genes for DLD1 and 1001 genes for SW480 cells).

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Colon Cancer

SUBMITTER: Michelle Palmieri  

LAB HEAD: Oliver Sieber

PROVIDER: PXD021936 | Pride | 2023-05-10

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
140606_michellep_16.raw Raw
140606_michellep_17.raw Raw
140606_michellep_18.raw Raw
140606_michellep_19.raw Raw
140606_michellep_20.raw Raw
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Publications

PI3Kα Translocation Mediates Nuclear PtdIns(3,4,5)P<sub>3</sub> Effector Signaling in Colorectal Cancer.

Palmieri Michelle M   Catimel Bruno B   Mouradov Dmitri D   Sakthianandeswaren Anuratha A   Kapp Eugene E   Ang Ching-Seng CS   Williamson Nicholas A NA   Nowell Cameron J CJ   Christie Michael M   Desai Jayesh J   Gibbs Peter P   Burgess Antony W AW   Sieber Oliver M OM  

Molecular & cellular proteomics : MCP 20230316 4


The canonical view of PI3Kα signaling describes phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P<sub>3</sub>) generation and activation of downstream effectors at the plasma membrane or at microtubule-bound endosomes. Here, we show that colorectal cancer (CRC) cell lines exhibit a diverse plasma membrane-nuclear distribution of PI3Kα, controlling corresponding levels of subcellular PtdIns(3,4,5)P<sub>3</sub> pools. PI3Kα nuclear translocation was mediated by the importin β-dependent nucl  ...[more]

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