Microtubule nucleation from the fibrous corona by LIC1-pericentrin promotes chromosome congression

Wu, Jingchao; Larreategui-Aparicio, Ainhoa; Lambers, Maaike L.A.; Bodor, Dani L.; Klaasen, Sjoerd J.; Tollenaar, Eveline A.P.; Ruijter-Villani, Marta de; Kops, Geert J.P.L.

doi:10.1016/j.cub.2023.01.010

Cited by 14 publications

(33 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast to control cells, which had abundant microtubules newly nucleated on kinetochores 3 minutes after washout of nocodazole, cells depleted of CENP-E showed virtually no microtubule nucleation on kinetochores (Figure 3A, B, S3A). Consistent with our prior observation that fibrous corona-derived microtubule nucleation requires the LIC1 subunit of dynein (Wu et al, 2023), depletion of CENP-E substantially removed LIC1 from kinetochores (Figure S3B, C). Inhibition of CENP-E motor activity by treatment with GSK923295 did not prevent kinetochore-derived microtubule nucleation (Figure S3D, E), and, consistently, neither did expression of the 2111-C fragment, which lacks the motor domain (Figure 3C, D).…”

Section: Farnesylated Cenp-e Is Essential For Kinetochore-derived Mic...supporting

confidence: 90%

“…Our prior work showed that fibrous coronas are the site of kinetochore-dependent microtubule nucleation, which aids spindle assembly and chromosome congression (Wu et al, 2023). To verify the requirement of CENP-E for fibrous corona expansion, we next assessed whether kinetochore-derived microtubule nucleation depends on CENP-E. To this end, we washed out nocodazole from RPE1 cells that had been treated with nocodazole overnight.…”

Section: Farnesylated Cenp-e Is Essential For Kinetochore-derived Mic...mentioning

confidence: 99%

“…RPE1 FlpIn cell line stably expressing EB3-TagRFPT-ires-GFP-ZWILCH was used previously (Wu et al, 2023). RPE1 FlpIn cell line inducible expressing LAP-SPINDLY FL and LAP-SPINDLY △N were used previously (Wu et al, 2023).…”

Section: Stable Cell Lines and Generationmentioning

confidence: 99%

“…Dynein/dynactin aid in microtubule capture and subsequent poleward transport of chromosomes along the microtubule lattice to facilitate eventual biorientation (Li et al, 2007;Savoian et al, 2000;Vorozhko et al, 2008;Yang et al, 2007). In addition, the LIC1 subunit of dynein promotes microtubule nucleation from the fibrous corona through interaction with the centrosomal protein pericentrin (Wu et al, 2023). Once kinetochores engage in end-on microtubule .…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A farnesyl-dependent structural role for CENP-E in expansion of the fibrous corona

Raas

Alcaraz

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Correct chromosome segregation during cell division depends on proper connections between spindle microtubules and kinetochores. During prometaphase, kinetochores are temporarily covered with a dense protein meshwork known as the fibrous corona. Formed by oligomerization of ROD/ZW10/ZWILCH-SPINDLY (RZZ-S) complexes, the fibrous corona promotes spindle assembly, chromosome orientation and spindle checkpoint signaling. The molecular requirements for formation of the fibrous corona are not fully understood. Here we show that the fibrous corona depends on the mitotic kinesin CENP-E, and that poorly expanded fibrous coronas after CENP-E depletion are functionally compromised. This previously unrecognized role for CENP-E does not require its motor activity but instead is driven by farnesyl modification of its C-terminal kinetochore- and microtubule-binding domain. We show that in cells CENP-E interacts with RZZ-S complexes in a farnesyl-dependent manner. CENP-E is recruited to kinetochores following RZZ-S, and - while not required for RZZ-S oligomerization per se - promotes subsequent fibrous corona expansion. Our comparative genomics analyses suggest that the farnesylation motif in CENP-E orthologs emerged alongside the full RZZ-S module in an ancestral lineage close to the fungi-animal split (Obazoa), revealing potential conservation of the mechanisms for fibrous corona formation. Our results show that proper spindle assembly has a potentially conserved non-motor contribution from the kinesin CENP-E through stabilization of the fibrous corona meshwork during its formation.

show abstract

Section: Farnesylated Cenp-e Is Essential For Kinetochore-derived Mic...supporting

confidence: 90%

Section: Farnesylated Cenp-e Is Essential For Kinetochore-derived Mic...mentioning

confidence: 99%

Section: Stable Cell Lines and Generationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A farnesyl-dependent structural role for CENP-E in expansion of the fibrous corona

Raas

Alcaraz

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition to promoting bi-orientation, the corona has also been shown to promote microtubule nucleation (Wu et al, 2023) and to contribute to the spindle assembly checkpoint (SAC), a pathway that delays anaphase onset until successful bi-orientation of all sister chromatid pairs (Fischer, 2022;Lara-Gonzalez et al, 2021). One of the main components of the SAC, the MAD1:MAD2 core complex, is also a constituent of the corona, where it also interacts with the CDK1:Cyclin B complex (Alfonso-Perez et al, 2019;Allan et al, 2020;Hoffman et al, 2001;Jackman et al, 2020).…”

mentioning

confidence: 99%

A mechanism that integrates microtubule motors of opposite polarity at the kinetochore corona

Cmentowski

d'Amico

Ciossani

et al. 2023

Preprint

View full text Add to dashboard Cite

Chromosome biorientation on the mitotic spindle is prerequisite to errorless genome inheritance. CENP-E (kinesin 7) and Dynein-Dynactin (DD), microtubule motors with opposite polarity, promote biorientation from the kinetochore corona, a polymeric structure whose assembly requires MPS1 kinase. The corona building block consists of ROD, Zwilch, ZW10, and the DD adaptor Spindly (RZZS). How CENP-E and DD are scaffolded and mutually coordinated in the corona remains unclear. Here, we report near-complete depletion of RZZS and DD from kinetochores after depletion of CENP-E and the outer kinetochore protein KNL1. With inhibited MPS1, CENP-E, which we show binds directly to RZZS, is required to retain kinetochore RZZS. An RZZS phosphomimetic mutant bypasses this requirement. With active MPS1, CENP-E is dispensable for corona expansion, but strictly required for physiological kinetochore accumulation of DD. Thus, we identify the corona as an integrated scaffold where CENP-E kinesin controls DD kinetochore loading for coordinated bidirectional transport of chromosome cargo.

show abstract

Cell biology: Kinetochores nucleate their own microtubules

Renda¹,

Khodjakov²

2023

Current Biology

View full text Add to dashboard Cite

Microtubule nucleation from the fibrous corona by LIC1-pericentrin promotes chromosome congression

Cited by 14 publications

References 87 publications

A farnesyl-dependent structural role for CENP-E in expansion of the fibrous corona

A farnesyl-dependent structural role for CENP-E in expansion of the fibrous corona

A mechanism that integrates microtubule motors of opposite polarity at the kinetochore corona

Cell biology: Kinetochores nucleate their own microtubules

Contact Info

Product

Resources

About