Analysis of the Conditions That Affect the Selective Processing of Endogenous Notch1 by ADAM10 and ADAM17

Rolake O. Alabi; Jose Lora; Arda B. Celen; Thorsten Maretzky; Carl P. Blobel

doi:10.3390/ijms22041846

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Analysis of the Conditions That Affect the Selective Processing of Endogenous Notch1 by ADAM10 and ADAM17

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Analysis of the Conditions That Affect the Selective Processing of Endogenous Notch1 by ADAM10 and ADAM17

Rolake O. Alabi, Jose Lora, Arda B. Celen, Thorsten Maretzky and Carl P. Blobel

International journal of molecular sciences, Vol.22(4), pp.1-17

02/01/2021

DOI: 10.3390/ijms22041846

PMCID: PMC7918056

PMID: 33673337

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Abstract

Notch signaling is critical for controlling a variety of cell fate decisions during metazoan development and homeostasis. This unique, highly conserved signaling pathway relies on cell-to-cell contact, which triggers the proteolytic release of the cytoplasmic domain of the membrane-anchored transcription factor Notch from the membrane. A disintegrin and metalloproteinase (ADAM) proteins are crucial for Notch activation by processing its S2 site. While ADAM10 cleaves Notch1 under physiological, ligand-dependent conditions, ADAM17 mainly cleaves Notch1 under ligand-independent conditions. However, the mechanism(s) that regulate the distinct contributions of these ADAMs in Notch processing remain unclear. Using cell-based assays in mouse embryonic fibroblasts (mEFs) lacking ADAM10 and/or ADAM17, we aimed to clarify what determines the relative contributions of ADAM10 and ADAM17 to ligand-dependent or ligand-independent Notch processing. We found that EDTA-stimulated ADAM17-dependent Notch1 processing is rapid and requires the ADAM17-regulators iRhom1 and iRhom2, whereas the Delta-like 4-induced ligand-dependent Notch1 processing is slower and requires ADAM10. The selectivity of ADAM17 for EDTA-induced Notch1 processing can most likely be explained by a preference for ADAM17 over ADAM10 for the Notch1 cleavage site and by the stronger inhibition of ADAM10 by EDTA. The physiological ADAM10-dependent processing of Notch1 cannot be compensated for by ADAM17 in Adam10-/- mEFs, or by other ADAMs shown here to be able to cleave the Notch1 cleavage site, such as ADAMs9, 12, and 19. Collectively, these results provide new insights into the mechanisms underlying the substrate selectivity of ADAM10 and ADAM17 towards Notch1.

Biochemistry & Molecular Biology

Chemistry

Chemistry, Multidisciplinary

Life Sciences & Biomedicine

Physical Sciences

Science & Technology

Details

Title: Subtitle: Analysis of the Conditions That Affect the Selective Processing of Endogenous Notch1 by ADAM10 and ADAM17
Creators: Rolake O. Alabi - Memorial Sloan Kettering Cancer Center
Jose Lora - Hospital for Special Surgery
Arda B. Celen - Hospital for Special Surgery
Thorsten Maretzky - Roy J. and Lucille A. Carver College of Medicine
Carl P. Blobel - Memorial Sloan Kettering Cancer Center
Resource Type: Journal article
Publication Details: International journal of molecular sciences, Vol.22(4), pp.1-17
DOI: 10.3390/ijms22041846
PMID: 33673337
PMCID: PMC7918056
NLM abbreviation: Int J Mol Sci
ISSN: 1661-6596
eISSN: 1422-0067
Publisher: Mdpi
Number of pages: 16
Grant note: R01 GM64750; R35 GM134907 / NIGMS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) T32 GM007739; T32EY00713820; T32HD606006; F31 GM136144; T32 GM008539 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
Language: English
Date published: 02/01/2021
Academic Unit: Internal Medicine
Record Identifier: 9984359923202771

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