Methylated histones on mitotic chromosomes promote topoisomerase IIα function for high fidelity chromosome segregation.
DNA Topoisomerase IIα (TopoIIα) decatenates sister chromatids, allowing their segregation in mitosis. Without the TopoIIα Strand Passage Reaction (SPR), chromosome bridges and ultra-fine DNA bridges (UFBs) arise in anaphase. The TopoIIα C-terminal domain is dispensable for the SPR in vitro but essential for mitotic functions in vivo. Here, we present evidence that the Chromatin Tether (ChT) within the CTD interacts with specific methylated nucleosomes and is crucial for high-fidelity chromosome segregation. Mutation of individual αChT residues disrupts αChT-nucleosome interaction, induces loss of segregation fidelity and reduces association of TopoIIα with chromosomes. Specific methyltransferase inhibitors reducing histone H3 or H4 methylation decreased TopoIIα at centromeres and increased segregation errors. Methyltransferase inhibition did not further increase aberrant anaphases in the ChT mutants, indicating a functional connection. The evidence reveals novel cellular regulation whereby TopoIIα specifically interacts with methylated nucleosomes via the αChT to ensure high-fidelity chromosome segregation.
Pubmed ID: 37197327 RIS Download
Research resources used in this publication
Additional research tools detected in this publication
Antibodies used in this publication
- IRDye 680RD Goat anti-Mouse IgG (RRID:AB_10956588)
- IRDye 680RD Goat anti-Rabbit IgG (RRID:AB_10956166)
- IRDye 800CW Goat anti-Mouse IgG (RRID:AB_621842)
- IRDye 800CW Goat anti-Rabbit IgG (RRID:AB_621843)
- Goat Anti-Guinea Pig IgG (H+L) Highly Cross-adsorbed Antibody, Alexa Fluor 647 Conjugated (RRID:AB_141882)
- Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 568 (RRID:AB_2534121)
- Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (RRID:AB_2576217)
- Anti-Human ANA-Centromere CREST-directed antibody Antibody, Unconjugated (RRID:AB_1282595)
- RFP antibody [5F8] (RRID:AB_2336064)
- Rabbit Anti-Histone H3, trimethyl (Lys27) Polyclonal antibody, Unconjugated (RRID:AB_310624)
- anti-CENP-C (RRID:AB_10693556)
- Mouse Anti-Histone H2B Monoclonal Antibody, Unconjugated, Clone 53H3 (RRID:AB_2295301)
- Histone H2A (D6O3A) Antibody (RRID:AB_2687875)
- Anti-Histone H3, phospho (Ser28) Antibody, Unconjugated (RRID:AB_823532)
- Histone H3K4me3 antibody (mAb) (RRID:AB_2793611)
- Histone H3K9me3 Antibody (RRID:AB_2687870)
- Mouse Anti-Histone H3 Monoclonal Antibody, Unconjugated, Clone 96C10 (RRID:AB_1642229)
- H3K27me3-mouse,H3K27me3-human (RRID:AB_2614987)
- H3K27me2-dmelanogaster (RRID:AB_448222)
- H3K27me3-human (RRID:AB_2616029)
- Histone H4K20me3 antibody (mAb) (RRID:AB_2650526)
- Monoclonal Anti-beta-Tubulin antibody produced in mouse (RRID:AB_477577)
- Histone H4 antibody (mAb) (RRID:AB_2793667)
- mCherry, rabbit polyclonal, Cat# RPCA-mCherry (RRID:AB_2571870)
- Rat Anti-DYKDDDDK Tag Monoclonal Antibody, Unconjugated (RRID:AB_10597743)
- Monoclonal ANTI-FLAG® M2 antibody produced in mouse (RRID:AB_262044)
Associated grants
- Agency: NIGMS NIH HHS, United States
Id: R01 GM130858
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This is a list of tools and resources that we have found mentioned in this publication.
DLD-1 (tool)
RRID:CVCL_0248
Cell line DLD-1 is a Cancer cell line with a species of origin Homo sapiens (Human)
View all literature mentionsIRDye 680RD Goat anti-Mouse IgG (antibody)
RRID:AB_10956588
This polyclonal targets IgG
View all literature mentionsIRDye 680RD Goat anti-Rabbit IgG (antibody)
RRID:AB_10956166
This polyclonal targets IgG
View all literature mentionsIRDye 800CW Goat anti-Mouse IgG (antibody)
RRID:AB_621842
This unknown targets IgG
View all literature mentionsIRDye 800CW Goat anti-Rabbit IgG (antibody)
RRID:AB_621843
This polyclonal secondary targets IgG
View all literature mentionsGoat Anti-Guinea Pig IgG (H+L) Highly Cross-adsorbed Antibody, Alexa Fluor 647 Conjugated (antibody)
RRID:AB_141882
This polyclonal secondary targets IgG (H+L)
View all literature mentionsGoat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 568 (antibody)
RRID:AB_2534121
This polyclonal secondary targets IgG (H+L)
View all literature mentionsGoat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (antibody)
RRID:AB_2576217
This polyclonal secondary targets IgG (H+L)
View all literature mentionsAnti-Human ANA-Centromere CREST-directed antibody Antibody, Unconjugated (antibody)
RRID:AB_1282595
This unknown targets Human ANA-Centromere CREST-directed antibody
View all literature mentionsRFP antibody [5F8] (antibody)
RRID:AB_2336064
This monoclonal targets dsRed, mRFP, mCherry , mPlum, mRFPruby
View all literature mentionsRabbit Anti-Histone H3, trimethyl (Lys27) Polyclonal antibody, Unconjugated (antibody)
RRID:AB_310624
This polyclonal targets Histone H3, trimethyl (Lys27)
View all literature mentionsanti-CENP-C (antibody)
RRID:AB_10693556
This unknown targets anti-CENP-C
View all literature mentionsMouse Anti-Histone H2B Monoclonal Antibody, Unconjugated, Clone 53H3 (antibody)
RRID:AB_2295301
This monoclonal targets Histone H2B
View all literature mentionsHistone H2A (D6O3A) Antibody (antibody)
RRID:AB_2687875
This monoclonal targets Histone H2A
View all literature mentionsAnti-Histone H3, phospho (Ser28) Antibody, Unconjugated (antibody)
RRID:AB_823532
This unknown targets Histone H3, phospho (Ser28)
View all literature mentionsHistone H3K4me3 antibody (mAb) (antibody)
RRID:AB_2793611
This monoclonal targets Histone H3K4me3
View all literature mentionsHistone H3K9me3 Antibody (antibody)
RRID:AB_2687870
This monoclonal targets H3K9me3
View all literature mentionsMouse Anti-Histone H3 Monoclonal Antibody, Unconjugated, Clone 96C10 (antibody)
RRID:AB_1642229
This monoclonal targets Histone H3
View all literature mentionsH3K27me3-mouse,H3K27me3-human (antibody)
RRID:AB_2614987
This unknown targets H3K27me3
View all literature mentionsH3K27me2-dmelanogaster (antibody)
RRID:AB_448222
This polyclonal targets H3K27me2
View all literature mentionsH3K27me3-human (antibody)
RRID:AB_2616029
This unknown targets H3K27me3
View all literature mentionsHistone H4K20me3 antibody (mAb) (antibody)
RRID:AB_2650526
This monoclonal targets Histone H4K20me3
View all literature mentionsMonoclonal Anti-beta-Tubulin antibody produced in mouse (antibody)
RRID:AB_477577
This monoclonal targets beta-Tubulin
View all literature mentionsHistone H4 antibody (mAb) (antibody)
RRID:AB_2793667
This monoclonal targets Histone H4
View all literature mentionsmCherry, rabbit polyclonal, Cat# RPCA-mCherry (antibody)
RRID:AB_2571870
This polyclonal targets
View all literature mentionsRat Anti-DYKDDDDK Tag Monoclonal Antibody, Unconjugated (antibody)
RRID:AB_10597743
This monoclonal targets Rat DYKDDDDK Tag
View all literature mentionsMonoclonal ANTI-FLAG® M2 antibody produced in mouse (antibody)
RRID:AB_262044
This monoclonal targets FLAG
View all literature mentions
