Global Interactome Mapping Reveals Pro-tumorigenic Interactions of NF-κB in Breast Cancer

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F23%3A00132137" target="_blank" >RIV/00216224:14310/23:00132137 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Global Interactome Mapping Reveals Pro-tumorigenic Interactions of NF-κB in Breast Cancer

Popis výsledku v původním jazyce

Introduction: NF-κB pathway plays a key role in immune response and inflammation, however, our previous results supported by data from other studies show its role also in cancer development and progression, including lymph node metastasis of luminal A breast cancer [1]. Here we used size exclusion chromatography (SEC) fractionation and protein correlation profiling (PCP) [2] to study the impact of NF-κB modulation on global protein interactome dynamics in luminal A breast cancer model. Methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of 160 SEC fractions of native MCF-7 lysates with inhibited or innate NF-κB activity was performed and the PrInCE algorithm [3] was applied for protein interaction mapping. AlphaPulldown methodology was employed for protein complex prediction. Immunoprecipitation with LC-MS/MS detection was used for characterization of NF-κB factor RELA interactome. Results: The co-fractionation experiment led to identification of 5460 protein groups in total (FDR = 0.01) and to detection of 7568 interactions among 1520 protein groups. Of these, 2564 interactions have been validated in independent datasets. NF-κB modulation was associated with rearrangement of protein complexes involved in NF-κB signaling and immune response, cell cycle regulation and DNA replication. Central NF-κB transcription factor RELA co-eluted with interactors of NF-κB activator PRMT5, both established and new complexes were confirmed by AlphaPulldown prediction. A complementary immunoprecipitation experiment recapitulated RELA interactions with other NF-κB factors, and associated NF -κB inhibition with decreased binding of NF-κB activators to RELA. Conclusions: This study describes an extensive network of pro-tumorigenic NF-κB interactions and its rearrangement in breast cancer that may have a therapeutic implications in tumors with high NF-κB activity. References: 1. Bouchal P. et al.: Mol Cell Proteomics, 14(7):1814-30. (2015) 2. Kristensen A.R. et al.: Nat Methods, 9(9):907-9. (2012) 3. Stacey R.G. et al.: BMC Bioinformatics, 23;18(1):457 (2017)

Název v anglickém jazyce

Global Interactome Mapping Reveals Pro-tumorigenic Interactions of NF-κB in Breast Cancer

Popis výsledku anglicky

Introduction: NF-κB pathway plays a key role in immune response and inflammation, however, our previous results supported by data from other studies show its role also in cancer development and progression, including lymph node metastasis of luminal A breast cancer [1]. Here we used size exclusion chromatography (SEC) fractionation and protein correlation profiling (PCP) [2] to study the impact of NF-κB modulation on global protein interactome dynamics in luminal A breast cancer model. Methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of 160 SEC fractions of native MCF-7 lysates with inhibited or innate NF-κB activity was performed and the PrInCE algorithm [3] was applied for protein interaction mapping. AlphaPulldown methodology was employed for protein complex prediction. Immunoprecipitation with LC-MS/MS detection was used for characterization of NF-κB factor RELA interactome. Results: The co-fractionation experiment led to identification of 5460 protein groups in total (FDR = 0.01) and to detection of 7568 interactions among 1520 protein groups. Of these, 2564 interactions have been validated in independent datasets. NF-κB modulation was associated with rearrangement of protein complexes involved in NF-κB signaling and immune response, cell cycle regulation and DNA replication. Central NF-κB transcription factor RELA co-eluted with interactors of NF-κB activator PRMT5, both established and new complexes were confirmed by AlphaPulldown prediction. A complementary immunoprecipitation experiment recapitulated RELA interactions with other NF-κB factors, and associated NF -κB inhibition with decreased binding of NF-κB activators to RELA. Conclusions: This study describes an extensive network of pro-tumorigenic NF-κB interactions and its rearrangement in breast cancer that may have a therapeutic implications in tumors with high NF-κB activity. References: 1. Bouchal P. et al.: Mol Cell Proteomics, 14(7):1814-30. (2015) 2. Kristensen A.R. et al.: Nat Methods, 9(9):907-9. (2012) 3. Stacey R.G. et al.: BMC Bioinformatics, 23;18(1):457 (2017)

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LX22NPO5102" target="_blank" >LX22NPO5102: Národní ústav pro výzkum rakoviny</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů