Identification of the Bok Interactome Using Proximity Labeling
The function of the Bcl-2 family member Bok is currently enigmatic, with various disparate roles reported, including mediation of apoptosis, regulation of mitochondrial morphology, binding to inositol 1,4,5-trisphosphate receptors, and regulation of uridine metabolism. To better define the roles of...
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Format: | Article |
Language: | English |
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Frontiers Media S.A.
2021-05-01
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Series: | Frontiers in Cell and Developmental Biology |
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Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2021.689951/full |
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author | Laura M. Szczesniak Caden G. Bonzerato Richard J. H. Wojcikiewicz |
author_facet | Laura M. Szczesniak Caden G. Bonzerato Richard J. H. Wojcikiewicz |
author_sort | Laura M. Szczesniak |
collection | DOAJ |
description | The function of the Bcl-2 family member Bok is currently enigmatic, with various disparate roles reported, including mediation of apoptosis, regulation of mitochondrial morphology, binding to inositol 1,4,5-trisphosphate receptors, and regulation of uridine metabolism. To better define the roles of Bok, we examined its interactome using TurboID-mediated proximity labeling in HeLa cells, in which Bok knock-out leads to mitochondrial fragmentation and Bok overexpression leads to apoptosis. Labeling with TurboID-Bok revealed that Bok was proximal to a wide array of proteins, particularly those involved in mitochondrial fission (e.g., Drp1), endoplasmic reticulum-plasma membrane junctions (e.g., Stim1), and surprisingly among the Bcl-2 family members, just Mcl-1. Comparison with TurboID-Mcl-1 and TurboID-Bak revealed that the three Bcl-2 family member interactomes were largely independent, but with some overlap that likely identifies key interactors. Interestingly, when overexpressed, Mcl-1 and Bok interact physically and functionally, in a manner that depends upon the transmembrane domain of Bok. Overall, this work shows that the Bok interactome is different from those of Mcl-1 and Bak, identifies novel proximities and potential interaction points for Bcl-2 family members, and suggests that Bok may regulate mitochondrial fission via Mcl-1 and Drp1. |
first_indexed | 2024-12-21T23:46:15Z |
format | Article |
id | doaj.art-c22271909a5b4c88bafee70afb25fbd5 |
institution | Directory Open Access Journal |
issn | 2296-634X |
language | English |
last_indexed | 2024-12-21T23:46:15Z |
publishDate | 2021-05-01 |
publisher | Frontiers Media S.A. |
record_format | Article |
series | Frontiers in Cell and Developmental Biology |
spelling | doaj.art-c22271909a5b4c88bafee70afb25fbd52022-12-21T18:46:06ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2021-05-01910.3389/fcell.2021.689951689951Identification of the Bok Interactome Using Proximity LabelingLaura M. SzczesniakCaden G. BonzeratoRichard J. H. WojcikiewiczThe function of the Bcl-2 family member Bok is currently enigmatic, with various disparate roles reported, including mediation of apoptosis, regulation of mitochondrial morphology, binding to inositol 1,4,5-trisphosphate receptors, and regulation of uridine metabolism. To better define the roles of Bok, we examined its interactome using TurboID-mediated proximity labeling in HeLa cells, in which Bok knock-out leads to mitochondrial fragmentation and Bok overexpression leads to apoptosis. Labeling with TurboID-Bok revealed that Bok was proximal to a wide array of proteins, particularly those involved in mitochondrial fission (e.g., Drp1), endoplasmic reticulum-plasma membrane junctions (e.g., Stim1), and surprisingly among the Bcl-2 family members, just Mcl-1. Comparison with TurboID-Mcl-1 and TurboID-Bak revealed that the three Bcl-2 family member interactomes were largely independent, but with some overlap that likely identifies key interactors. Interestingly, when overexpressed, Mcl-1 and Bok interact physically and functionally, in a manner that depends upon the transmembrane domain of Bok. Overall, this work shows that the Bok interactome is different from those of Mcl-1 and Bak, identifies novel proximities and potential interaction points for Bcl-2 family members, and suggests that Bok may regulate mitochondrial fission via Mcl-1 and Drp1.https://www.frontiersin.org/articles/10.3389/fcell.2021.689951/fullBcl-2 related ovarian killerB-cell lymphoma 2 (Bcl-2) familyproximity labelingmyeloid-cell leukemia 1apoptosis |
spellingShingle | Laura M. Szczesniak Caden G. Bonzerato Richard J. H. Wojcikiewicz Identification of the Bok Interactome Using Proximity Labeling Frontiers in Cell and Developmental Biology Bcl-2 related ovarian killer B-cell lymphoma 2 (Bcl-2) family proximity labeling myeloid-cell leukemia 1 apoptosis |
title | Identification of the Bok Interactome Using Proximity Labeling |
title_full | Identification of the Bok Interactome Using Proximity Labeling |
title_fullStr | Identification of the Bok Interactome Using Proximity Labeling |
title_full_unstemmed | Identification of the Bok Interactome Using Proximity Labeling |
title_short | Identification of the Bok Interactome Using Proximity Labeling |
title_sort | identification of the bok interactome using proximity labeling |
topic | Bcl-2 related ovarian killer B-cell lymphoma 2 (Bcl-2) family proximity labeling myeloid-cell leukemia 1 apoptosis |
url | https://www.frontiersin.org/articles/10.3389/fcell.2021.689951/full |
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