Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C

Dimitriya H Garvanska; Marie Sofie Yoo Larsen; Jakob Nilsson

doi:10.1242/bio.020842

Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C

Dimitriya H Garvanska, Marie Sofie Yoo Larsen, Jakob Nilsson

4 Citationer (Scopus)

Abstract

The spindle assembly checkpoint (SAC) inhibits the anaphase-promoting complex/cyclosome (APC/C) in response to unattached kinetochores by generating a diffusible inhibitor termed the mitotic checkpoint complex (MCC). At metaphase, rapid activation of the APC/C requires removal of the MCC, a process that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor contribution to SAC silencing in HCT116 cells. Strikingly in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E2 enzyme UBE2D. In conclusion, we provide in vivo insight into the APC/C E2 module and its interplay with SAC silencing components.

Originalsprog	Engelsk
Tidsskrift	Biology Open
Vol/bind	5
Sider (fra-til)	1441-1448
ISSN	2046-6390
DOI	https://doi.org/10.1242/bio.020842
Status	Udgivet - 15 okt. 2016

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10.1242/bio.020842Licens: CC BY

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title = "Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C",

abstract = "The spindle assembly checkpoint (SAC) inhibits the anaphase-promoting complex/cyclosome (APC/C) in response to unattached kinetochores by generating a diffusible inhibitor termed the mitotic checkpoint complex (MCC). At metaphase, rapid activation of the APC/C requires removal of the MCC, a process that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor contribution to SAC silencing in HCT116 cells. Strikingly in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E2 enzyme UBE2D. In conclusion, we provide in vivo insight into the APC/C E2 module and its interplay with SAC silencing components.",

author = "Garvanska, {Dimitriya H} and Larsen, {Marie Sofie Yoo} and Jakob Nilsson",

note = "{\textcopyright} 2016. Published by The Company of Biologists Ltd.",

year = "2016",

month = oct,

day = "15",

doi = "10.1242/bio.020842",

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TY - JOUR

T1 - Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C

AU - Garvanska, Dimitriya H

AU - Larsen, Marie Sofie Yoo

AU - Nilsson, Jakob

PY - 2016/10/15

Y1 - 2016/10/15

N2 - The spindle assembly checkpoint (SAC) inhibits the anaphase-promoting complex/cyclosome (APC/C) in response to unattached kinetochores by generating a diffusible inhibitor termed the mitotic checkpoint complex (MCC). At metaphase, rapid activation of the APC/C requires removal of the MCC, a process that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor contribution to SAC silencing in HCT116 cells. Strikingly in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E2 enzyme UBE2D. In conclusion, we provide in vivo insight into the APC/C E2 module and its interplay with SAC silencing components.

AB - The spindle assembly checkpoint (SAC) inhibits the anaphase-promoting complex/cyclosome (APC/C) in response to unattached kinetochores by generating a diffusible inhibitor termed the mitotic checkpoint complex (MCC). At metaphase, rapid activation of the APC/C requires removal of the MCC, a process that has been shown to depend on the APC/C E2 enzymes, UBE2C and UBE2S. Here we investigate the in vivo role of the APC/C E2 enzymes in SAC silencing using CRISPR/Cas9 genetically engineered HCT116 UBE2C or UBE2S null cell lines. Using live cell assays, we show that UBE2C and UBE2S make a minor contribution to SAC silencing in HCT116 cells. Strikingly in cells specifically lacking UBE2C, we observe a strong synergistic inhibition of mitotic progression when we stabilize the MCC on the APC/C by depleting APC15, potentially reflecting increased competition between the MCC and the remaining initiating E2 enzyme UBE2D. In conclusion, we provide in vivo insight into the APC/C E2 module and its interplay with SAC silencing components.

U2 - 10.1242/bio.020842

DO - 10.1242/bio.020842

M3 - Journal article

C2 - 27591192

SN - 2046-6390

VL - 5

SP - 1441

EP - 1448

JO - Biology Open

JF - Biology Open

ER -

Synergistic inhibition of the APC/C by the removal of APC15 in HCT116 cells lacking UBE2C

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