Identification of Endogenous Adenomatous Polyposis Coli Interaction Partners and β-Catenin-Independent Targets by Proteomics

Olesja Popow, Joao A. Paulo, Michael Tatham, Melanie Volk, Alejandro Rojas-Fernandez, Nicolas Loyer, Ian Newton, Jens Januschke, Kevin M. Haigis, Inke Nathke (Lead / Corresponding author)

Research output: Contribution to journalArticle

Abstract

Adenomatous Polyposis Coli (APC) is the most frequently mutated gene in colorectal cancer. APC negatively regulates the Wnt signaling pathway by promoting the degradation of β-catenin, but the extent to which APC exerts Wnt/β-catenin-independent tumor-suppressive activity is unclear. To identify interaction partners and β-catenin-independent targets of endogenous, full-length APC, we applied label-free and multiplexed tandem mass tag-based mass spectrometry. Affinity enrichment-mass spectrometry identified more than 150 previously unidentified APC interaction partners. Moreover, our global proteomic analysis revealed that roughly half of the protein expression changes that occur in response to APC loss are independent of β-catenin. Combining these two analyses, we identified Misshapen-like kinase 1 (MINK1) as a putative substrate of an APC-containing destruction complex. We validated the interaction between endogenous MINK1 and APC and further confirmed the negative, and β-catenin-independent, regulation of MINK1 by APC. Increased Mink1/Msn levels were also observed in mouse intestinal tissue and Drosophila follicular cells expressing mutant Apc/APC when compared with wild-type tissue/cells. Collectively, our results highlight the extent and importance of Wnt-independent APC functions in epithelial biology and disease. IMPLICATIONS: The tumor-suppressive function of APC, the most frequently mutated gene in colorectal cancer, is mainly attributed to its role in β-catenin/Wnt signaling. Our study substantially expands the list of APC interaction partners and reveals that approximately half of the changes in the cellular proteome induced by loss of APC function are mediated by β-catenin-independent mechanisms.

Original languageEnglish
Pages (from-to)1828-1841
Number of pages14
JournalMolecular Cancer Research
Volume17
Issue number9
Early online date3 Jun 2019
DOIs
Publication statusPublished - Sep 2019

Fingerprint

Catenins
Adenomatous Polyposis Coli
Proteomics
Phosphotransferases
Colorectal Neoplasms
Mass Spectrometry
Wnt Signaling Pathway
Proteome
Genes
Drosophila

Keywords

  • Adenomatous polyposis coli
  • destruction complex
  • colorectal cancer
  • proteomics
  • Misshapen-like kinase 1

Cite this

@article{63e768cb81e0489a9bf69854ed16d291,
title = "Identification of Endogenous Adenomatous Polyposis Coli Interaction Partners and β-Catenin-Independent Targets by Proteomics",
abstract = "Adenomatous Polyposis Coli (APC) is the most frequently mutated gene in colorectal cancer. APC negatively regulates the Wnt signaling pathway by promoting the degradation of β-catenin, but the extent to which APC exerts Wnt/β-catenin-independent tumor-suppressive activity is unclear. To identify interaction partners and β-catenin-independent targets of endogenous, full-length APC, we applied label-free and multiplexed tandem mass tag-based mass spectrometry. Affinity enrichment-mass spectrometry identified more than 150 previously unidentified APC interaction partners. Moreover, our global proteomic analysis revealed that roughly half of the protein expression changes that occur in response to APC loss are independent of β-catenin. Combining these two analyses, we identified Misshapen-like kinase 1 (MINK1) as a putative substrate of an APC-containing destruction complex. We validated the interaction between endogenous MINK1 and APC and further confirmed the negative, and β-catenin-independent, regulation of MINK1 by APC. Increased Mink1/Msn levels were also observed in mouse intestinal tissue and Drosophila follicular cells expressing mutant Apc/APC when compared with wild-type tissue/cells. Collectively, our results highlight the extent and importance of Wnt-independent APC functions in epithelial biology and disease. IMPLICATIONS: The tumor-suppressive function of APC, the most frequently mutated gene in colorectal cancer, is mainly attributed to its role in β-catenin/Wnt signaling. Our study substantially expands the list of APC interaction partners and reveals that approximately half of the changes in the cellular proteome induced by loss of APC function are mediated by β-catenin-independent mechanisms.",
keywords = "Adenomatous polyposis coli, destruction complex, colorectal cancer, proteomics, Misshapen-like kinase 1",
author = "Olesja Popow and Paulo, {Joao A.} and Michael Tatham and Melanie Volk and Alejandro Rojas-Fernandez and Nicolas Loyer and Ian Newton and Jens Januschke and Haigis, {Kevin M.} and Inke Nathke",
note = "Olesja Popow - CRUK PhD fellowship, Jo{\~a}o A. Paulo - NIH/NIDDK grant K01 DK098285, Michael H. Tatham – supported by CRUK program grant to R. Hay, Alejandro Rojas-Fernandez - FONDECYT 11150532 and PAI79150075, Nicolas Loyer and Jens Januschke - Sir Henry Dale fellowship (Wellcome/Royal Society: 100031Z/12/Z), Kevin M. Haigis - NIH/NCI grant U01CA199252, Ian P. Newton and Inke N{\"a}thke – CRUK program grant to Inke N{\"a}thke.",
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Identification of Endogenous Adenomatous Polyposis Coli Interaction Partners and β-Catenin-Independent Targets by Proteomics. / Popow, Olesja; Paulo, Joao A.; Tatham, Michael; Volk, Melanie; Rojas-Fernandez, Alejandro; Loyer, Nicolas; Newton, Ian; Januschke, Jens; Haigis, Kevin M.; Nathke, Inke (Lead / Corresponding author).

In: Molecular Cancer Research, Vol. 17, No. 9, 09.2019, p. 1828-1841.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of Endogenous Adenomatous Polyposis Coli Interaction Partners and β-Catenin-Independent Targets by Proteomics

AU - Popow, Olesja

AU - Paulo, Joao A.

AU - Tatham, Michael

AU - Volk, Melanie

AU - Rojas-Fernandez, Alejandro

AU - Loyer, Nicolas

AU - Newton, Ian

AU - Januschke, Jens

AU - Haigis, Kevin M.

AU - Nathke, Inke

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