The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface

McKinley, Kara L.; Sekulić, Nikolina; Guo, Lucie Y.; Tsinman, Tonia; Black, Ben E.; Cheeseman, Iain M.

doi:10.1016/j.molcel.2015.10.027

Cited by 162 publications

(284 citation statements)

References 62 publications

Supporting

Mentioning

263

Contrasting

Order By: Relevance

“…At 3 d (CENP-W) or 5 d (CENP-C or CENP-T) after doxycycline induction, chromatinassociation and c-CO-FISH pattern were monitored. As previously reported (35), all knockout treatments led to decrease of chromatinor centromere-associated CENP-T without affecting CENP-A, whereas CENP-C knockout, but not CENP-T or CENP-W knockout, decreased chromatin-associated CENP-C (SI Appendix, Fig. S7 D and E).…”

Section: Superresolution Microscopy Of Centromere Organization Revealssupporting

confidence: 83%

See 1 more Smart Citation

Integrity of the human centromere DNA repeats is protected by CENP-A, CENP-C, and CENP-T

Giunta

Funabiki

2017

Proc. Natl. Acad. Sci. U.S.A.

105

View full text Add to dashboard Cite

Centromeres are highly specialized chromatin domains that enable chromosome segregation and orchestrate faithful cell division. Human centromeres are composed of tandem arrays of α-satellite DNA, which spans up to several megabases. Little is known about the mechanisms that maintain integrity of the long arrays of α-satellite DNA repeats. Here, we monitored centromeric repeat stability in human cells using chromosome-orientation fluorescent in situ hybridization (CO-FISH). This assay detected aberrant centromeric CO-FISH patterns consistent with sister chromatid exchange at the frequency of 5% in primary tissue culture cells, whereas higher levels were seen in several cancer cell lines and during replicative senescence. To understand the mechanism(s) that maintains centromere integrity, we examined the contribution of the centromere-specific histone variant CENP-A and members of the constitutive centromere-associated network (CCAN), CENP-C, CENP-T, and CENP-W. Depletion of CENP-A and CCAN proteins led to an increase in centromere aberrations, whereas enhancing chromosome missegregation by alternative methods did not, suggesting that CENP-A and CCAN proteins help maintain centromere integrity independently of their role in chromosome segregation. Furthermore, superresolution imaging of centromeric CO-FISH using structured illumination microscopy implied that CENP-A protects α-satellite repeats from extensive rearrangements. Our study points toward the presence of a centromere-specific mechanism that actively maintains α-satellite repeat integrity during human cell proliferation.centromere | genome integrity | α-satellite repeats | cancer | CO-FISH

show abstract

Section: Superresolution Microscopy Of Centromere Organization Revealssupporting

confidence: 83%

“…To verify these results, we used HeLa cell lines where CENP-C, CENP-T, and CENP-W can be conditionally knocked out by doxycycline-mediated induction of Cas9 (35). At 3 d (CENP-W) or 5 d (CENP-C or CENP-T) after doxycycline induction, chromatinassociation and c-CO-FISH pattern were monitored.…”

Section: Superresolution Microscopy Of Centromere Organization Revealsmentioning

confidence: 99%

Integrity of the human centromere DNA repeats is protected by CENP-A, CENP-C, and CENP-T

Giunta

Funabiki

2017

Proc. Natl. Acad. Sci. U.S.A.

105

View full text Add to dashboard Cite

show abstract

“…Studies in Saccharomyces cerevisiae report a role for the COMA complex (CENP-O/-P/-Q/-R/-U complex homolog) in the looping of centromere chromatin (45). However, our results plus other recent studies have failed to identify a function for this complex in vertebrate kinetochores (37,41).…”

Section: The Role Of Ccan Proteins In the Maintenance Of Kinetochorecontrasting

confidence: 57%

“…In contrast, CENP-C has a web of interactions with other CCAN members, including CENP-A and CENP-H/-I/-K/-M (35,36,41) as well as outer kinetochore components (10,42). CENP-T/W/S/X also interacts both with CENP-H/-I/-K/-M (10) and the Ndc80 complex (13,43).…”

Section: The Role Of Ccan Proteins In the Maintenance Of Kinetochorementioning

confidence: 99%

Stepwise unfolding supports a subunit model for vertebrate kinetochores

Vargiu

Макаров

Allan

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

During cell division, interactions between microtubules and chromosomes are mediated by the kinetochore, a proteinaceous structure located at the primary constriction of chromosomes. In addition to the centromere histone centromere protein A (CENP-A), 15 other members of the constitutive centromere associated network (CCAN) participate in the formation of a chromatin-associated scaffold that supports kinetochore structure. We performed a targeted screen analyzing unfolded centrochromatin from CENP-depleted chromosomes. Our results revealed that CENP-C and CENP-S are critical for the stable folding of mitotic kinetochore chromatin. Multipeak fitting algorithms revealed the presence of an organized pattern of centrochromatin packing consistent with arrangement of CENP-A-containing nucleosomes into up to five chromatin "subunits"-each containing roughly 20-30 nucleosomes. These subunits could be either layers of a boustrophedon or small loops of centromeric chromatin.centromere | kinetochore | CENP-A | centrochromatin | boustrophedon

show abstract

“…To test this hypothesis, we made inducible CRISPR/Cas9 RPE1 cell lines to knock out NuMA (Figure 3-figure supplement 1) (McKinley et al, 2015;Wang et al, 2015). Indeed, when NuMA was knocked out -causing elongated, heterogeneously disorganized spindles with detached centrosomes -ablation-created minus-ends were no longer transported toward poles by dynein (Figure 3A-C; Video 5).…”

Section: Numa Localizes To Minus-ends Independently Of Dyneinmentioning

confidence: 99%

NuMA Recruits Dynein Activity to Microtubule Minus-Ends at Mitosis

Hueschen

Kenny

et al. 2018

Biophysical Journal

View full text Add to dashboard Cite

To build the spindle at mitosis, motors exert spatially regulated forces on microtubules. We know that dynein pulls on mammalian spindle microtubule minus-ends, and this localized activity at ends is predicted to allow dynein to cluster microtubules into poles. How dynein becomes enriched at minus-ends is not known. Here, we use quantitative imaging and laser ablation to show that NuMA targets dynactin to minus-ends, localizing dynein activity there. NuMA is recruited to new minus-ends independently of dynein and more quickly than dynactin; both NuMA and dynactin display specific, steady-state binding at minus-ends. NuMA localization to minus-ends involves a C-terminal region outside NuMA's canonical microtubule-binding domain and is independent of minus-end binders g-TuRC, CAMSAP1, and KANSL1/3. Both NuMA's minus-endbinding and dynein-dynactin-binding modules are required to rescue focused, bipolar spindle organization. Thus, NuMA may serve as a mitosis-specific minus-end cargo adaptor, targeting dynein activity to minus-ends to cluster spindle microtubules into poles.

show abstract

The CENP-L-N Complex Forms a Critical Node in an Integrated Meshwork of Interactions at the Centromere-Kinetochore Interface

Cited by 162 publications

References 62 publications

Integrity of the human centromere DNA repeats is protected by CENP-A, CENP-C, and CENP-T

Integrity of the human centromere DNA repeats is protected by CENP-A, CENP-C, and CENP-T

Stepwise unfolding supports a subunit model for vertebrate kinetochores

NuMA Recruits Dynein Activity to Microtubule Minus-Ends at Mitosis

Contact Info

Product

Resources

About