Mechanistic insight into TRIP13-catalyzed Mad2 structural transition and spindle checkpoint silencing

Brulotte, Melissa Lynn; Jeong, Byung Cheon; Li, Faxiang; Li, Bing; Yu, Eric B.; Wu, Qiong; Brautigam, Chad A.; Yu, Hongtao; Luo, Xuelian

doi:10.1038/s41467-017-02012-2

Cited by 42 publications

(35 citation statements)

References 78 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since both Mad2 and REV7 form a conformational dimer to recruit downstream effectors, this may be a universal mechanism utilized by HORMA proteins 26 . It is known that Mad2 is regulated by p31 comet and ATPase TRIP13 through structural remodeling 29,30 . However, how shieldin is regulated is unknown.…”

Section: Discussionmentioning

confidence: 99%

Molecular basis for assembly of the shieldin complex and its implications for NHEJ

et al. 2020

View full text Add to dashboard Cite

Shieldin, including SHLD1, SHLD2, SHLD3 and REV7, functions as a bridge linking 53BP1-RIF1 and single-strand DNA to suppress the DNA termini nucleolytic resection during nonhomologous end joining (NHEJ). However, the mechanism of shieldin assembly remains unclear. Here we present the crystal structure of the SHLD3-REV7-SHLD2 ternary complex and reveal an unexpected C (closed)-REV7-O (open)-REV7 conformational dimer mediated by SHLD3. We show that SHLD2 interacts with O-REV7 and the N-terminus of SHLD3 by forming β sheet sandwich. Disruption of the REV7 conformational dimer abolishes the assembly of shieldin and impairs NHEJ efficiency. The conserved FXPWFP motif of SHLD3 binds to C-REV7 and blocks its binding to REV1, which excludes shieldin from the REV1/Pol ζ translesion synthesis (TLS) complex. Our study reveals the molecular architecture of shieldin assembly, elucidates the structural basis of the REV7 conformational dimer, and provides mechanistic insight into orchestration between TLS and NHEJ.

show abstract

Section: Discussionmentioning

confidence: 99%

Molecular basis for assembly of the shieldin complex and its implications for NHEJ

et al. 2020

View full text Add to dashboard Cite

show abstract

“…5 ). Recently, Brulotte et al 66 showed that in vitro low levels of TRIP13 were unable to quickly reactivate APC/C Cdc20 that had been pre-incubated with MCC components, but were sufficient to disassemble-free MCC and prevent APC/C Cdc20 inhibition. These data clearly show that free MCC is the preferred substrate of TRIP13, but does not eliminate the possibility of low-level activity on APC/C-MCC complexes.…”

Section: Discussionmentioning

confidence: 99%

TRIP13 and APC15 drive mitotic exit by turnover of interphase- and unattached kinetochore-produced MCC

Kim

Han

et al. 2018

Nat Commun

View full text Add to dashboard Cite

The mitotic checkpoint ensures accurate chromosome segregation through assembly of the mitotic checkpoint complex (MCC), a soluble inhibitor of the anaphase-promoting complex/cyclosome (APC/C) produced by unattached kinetochores. MCC is also assembled during interphase by Mad1/Mad2 bound at nuclear pores, thereby preventing premature mitotic exit prior to kinetochore maturation and checkpoint activation. Using degron tagging to rapidly deplete the AAA+ ATPase TRIP13, we show that its catalytic activity is required to maintain a pool of open-state Mad2 for MCC assembly, thereby supporting mitotic checkpoint activation, but is also required for timely mitotic exit through catalytic disassembly of MCC. Strikingly, combining TRIP13 depletion with elimination of APC15-dependent Cdc20 ubiquitination/degradation results in a complete inability to exit mitosis, even when MCC assembly at unattached kinetochores is prevented. Thus, mitotic exit requires MCC produced either in interphase or mitosis to be disassembled by TRIP13-catalyzed removal of Mad2 or APC15-driven ubiquitination/degradation of its Cdc20 subunit.

show abstract

“…Cell pellets were washed 1x with PBS and flash frozen in liquid nitrogen. Ubiquitylation reactions were performed as previously described ( Brulotte et al, 2017 ; Ji et al, 2017 ). Briefly, APC/C was isolated from HeLa cell extracts with anti-APC3 antibody coupled Protein A beads (Bio-Rad).…”

Section: Methodsmentioning

confidence: 99%

Sumoylation promotes optimal APC/C activation and timely anaphase

Lee

et al. 2018

eLife

Self Cite

View full text Add to dashboard Cite

The Anaphase Promoting Complex/Cyclosome (APC/C) is a ubiquitin E3 ligase that functions as the gatekeeper to mitotic exit. APC/C activity is controlled by an interplay of multiple pathways during mitosis, including the spindle assembly checkpoint (SAC), that are not yet fully understood. Here, we show that sumoylation of the APC4 subunit of the APC/C peaks during mitosis and is critical for timely APC/C activation and anaphase onset. We have also identified a functionally important SUMO interacting motif in the cullin-homology domain of APC2 located near the APC4 sumoylation sites and APC/C catalytic core. Our findings provide evidence of an important regulatory role for SUMO modification and binding in affecting APC/C activation and mitotic exit.

show abstract

Mechanistic insight into TRIP13-catalyzed Mad2 structural transition and spindle checkpoint silencing

Cited by 42 publications

References 78 publications

Molecular basis for assembly of the shieldin complex and its implications for NHEJ

Molecular basis for assembly of the shieldin complex and its implications for NHEJ

TRIP13 and APC15 drive mitotic exit by turnover of interphase- and unattached kinetochore-produced MCC

Sumoylation promotes optimal APC/C activation and timely anaphase

Contact Info

Product

Resources

About