The effects of IFITM1 and IFITM3 gene deletion on IFNγ stimulated protein synthesis

Maria Gómez-Herranz , Marta Nekulova , Jakub Faktor , Lenka Hernychova , Sachin Kote , Elizabeth H. Sinclair , Rudolf Nenutil , Borivoj Vojtesek , Kathryn L. Ball , Ted R. Hupp

Abstract

Interferon-induced transmembrane proteins IFITM1 and IFITM3 (IFITM1/3) play a role in both RNA viral restriction and in human cancer progression. Using immunohistochemical staining of FFPE tissue, we identified subgroups of cervical cancer patients where IFITM1/3 protein expression is inversely related to metastasis. Guide RNA-CAS9 methods were used to develop an isogenic IFITM1/IFITM3 double null cervical cancer model in order to define dominant pathways triggered by presence or absence of IFITM1/3 signalling. A pulse SILAC methodology identified IRF1, HLA-B, and ISG15 as the most dominating IFNγ inducible proteins whose synthesis was attenuated in the IFITM1/IFITM3 double-null cells. Conversely, SWATH-IP mass spectrometry of ectopically expressed SBP-tagged IFITM1 identified ISG15 and HLA-B as dominant co-associated proteins. ISG15ylation was attenuated in IFNγ treated IFITM1/IFITM3 double-null cells. Proximity ligation assays indicated that HLA-B can interact with IFITM1/3 proteins in parental SiHa cells. Cell surface expression of HLA-B was attenuated in IFNγ treated IFITM1/IFITM3 double-null cells. SWATH-MS proteomic screens in cells treated with IFITM1-targeted siRNA cells resulted in the attenuation of an interferon regulated protein subpopulation including MHC Class I molecules as well as IFITM3, STAT1, B2M, and ISG15. These data have implications for the function of IFITM1/3 in mediating IFNγ stimulated protein synthesis including ISG15ylation and MHC Class I production in cancer cells. The data together suggest that pro-metastatic growth associated with IFITM1/3 negative cervical cancers relates to attenuated expression of MHC Class I molecules that would support tumor immune escape.
Author Maria Gómez-Herranz
Maria Gómez-Herranz,,
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, Marta Nekulova
Marta Nekulova,,
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, Jakub Faktor
Jakub Faktor,,
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, Lenka Hernychova
Lenka Hernychova,,
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, Sachin Kote (ICCVS)
Sachin Kote,,
- International Centre for Cancer Vaccine Science
, Elizabeth H. Sinclair
Elizabeth H. Sinclair,,
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, Rudolf Nenutil
Rudolf Nenutil,,
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, Borivoj Vojtesek
Borivoj Vojtesek,,
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, Kathryn L. Ball
Kathryn L. Ball,,
-
, Ted R. Hupp (ICCVS)
Ted R. Hupp,,
- International Centre for Cancer Vaccine Science
Journal seriesCellular Signalling, ISSN 0898-6568, (N/A 100 pkt)
Issue year2019
Vol60
Pages39-56
Publication size in sheets0.85
Keywords in EnglishCervical cancer, IFITM1, Interferon, SILAC mass spectrometry, MHC Class I molecule, CAS9 gene editing
ASJC Classification1307 Cell Biology
DOIDOI:10.1016/j.cellsig.2019.03.024
URL https://doi.org/10.1016/j.cellsig.2019.03.024
Languageen angielski
Score (nominal)100
Score sourcejournalList
ScoreMinisterial score = 100.0, 28-01-2020, ArticleFromJournal
Publication indicators Scopus SNIP (Source Normalised Impact per Paper): 2018 = 0.862; WoS Impact Factor: 2018 = 3.388 (2) - 2018=3.844 (5)
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