Dataset Information

IFITM3 amino acid mutants phosphoproteomics

ABSTRACT: To investigate the role of IFITM3 in B-ALL, patient derived xenograft cells PDX2 were transduced with either a the constitutively membrane associated IFITM3-Y20E mutant or EV control. Additional mutation of PIP3 binding sites within the conserved internal loop region - K83/K104 or R85/R87/K88 - were introduced to test the predicted function of these sites.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): B Cell, Cell Culture

DISEASE(S): Acute Leukemia

SUBMITTER: Mark Robinson

LAB HEAD: Markus Muschen

PROVIDER: PXD020696 | Pride | 2020-12-18

REPOSITORIES: Pride

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Publications

IFITM3 functions as a PIP3 scaffold to amplify PI3K signalling in B cells.

Lee Jaewoong J Robinson Mark E ME Ma Ning N Artadji Dewan D Ahmed Mohamed A MA Xiao Gang G Sadras Teresa T Deb Gauri G Winchester Janet J Cosgun Kadriye Nehir KN Geng Huimin H Chan Lai N LN Kume Kohei K Miettinen Teemu P TP Zhang Ye Y Nix Matthew A MA Klemm Lars L Chen Chun Wei CW Chen Jianjun J Khairnar Vishal V Wiita Arun P AP Thomas-Tikhonenko Andrei A Farzan Michael M Jung Jae U JU Weinstock David M DM Manalis Scott R SR Diamond Michael S MS Vaidehi Nagarajan N Müschen Markus M

Nature 20201104 7838

Interferon-induced transmembrane protein 3 (IFITM3) has previously been identified as an endosomal protein that blocks viral infection<sup>1-3</sup>. Here we studied clinical cohorts of patients with B cell leukaemia and lymphoma, and identified IFITM3 as a strong predictor of poor outcome. In normal resting B cells, IFITM3 was minimally expressed and mainly localized in endosomes. However, engagement of the B cell receptor (BCR) induced both expression of IFITM3 and phosphorylation of this prot ...[more]

PMID: 33149299

Similar Datasets

Project description:Interferon-induced transmembrane protein (IFITM1) plays a dual role in restriction of RNA viruses and in metastatic cancer cell growth. The signal transduction events that are orchestrated by IFITM1 are not well defined. We set out to identify IFITM1 interacting proteins to begin to define its mechanism of action. Affinity purification of SBP-tagged IFITM1, coupled to SWATH-mass spectrometry, identified significantly higher confidence interacting proteins from IFN- treated cells, relative to non-treated cells. This promoted an examination of the protein synthetic machinery in order to determine whether IFITM1 can impact on ribosomal proteome from IFN- treated cells. Isogenic ifitm1 null and ifitm1-ifitm3 null-SiHa cell panels were generated using CRISPR gRNAs to define signaling events that are linked to IFN--dependent protein synthesis. Ultracentrifugation sedimentation of cell lysates from interferon gamma treated wt and ifitm1-ifitm3 null-SiHa cells was carried out to isolate ribosomal constituents. Although SWATH-mass spectrometry of the 40S, 60S, and 80S fractions demonstrated changes in selected protein content, a significant reduction in A254 (RNA) in the 80S ribosomal fractions suggested a relatively select defect in 80S ribosomal biogenesis in ifitm1-ifitm3 null cells. The localization of IFITM1 to ribosomal protein components prompted an analysis of IFN--dependent protein synthesis using pulse SILAC. STAT1 and B2M were two dominant proteins whose synthesis increased equivalently in wt or ifitm1-ifitm3 null cells. However, MHC class I molecules and ISG15 were the most highly suppressed IFN--responsive proteins in the ifitm1-ifitm3 double null cells and this was confirmed using ifitm1 single null cells. Transient depletion of IFITM1 using targeted siRNA also depleted MHC class I molecules as well as IFITM3, STAT1, B2M, and ISG15. These data have implications for the function of IFITM1 in mediating IFN--stimulated protein synthesis and associated antigen presentation during either oncogenic or anti-viral signalling.

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