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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Main Authors: Laura M. Szczesniak, Caden G. Bonzerato, Richard J. H. Wojcikiewicz
Format: Article
Language:English
Published: Frontiers Media S.A. 2021-05-01
Series:Frontiers in Cell and Developmental Biology
Subjects:
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.
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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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