Molecular basis of outer kinetochore assembly on CENP-T

Veld, Pim J Huis in 't; Jeganathan, Sadasivam; Petrovic, Arsen; Singh, Priyanka; John, Juliane; Krenn, Veronica; Weissmann, Florian; Bange, Tanja; Musacchio, Andrea

doi:10.7554/elife.21007

Cited by 124 publications

(191 citation statements)

References 61 publications

(132 reference statements)

Supporting

Mentioning

161

Contrasting

Order By: Relevance

“…Previous low-resolution negative stain EM analyses depicted the Mis12 complex (known as MIND complex in S. cerevisiae ) as a ~20 nm rod [234,235,236,237]. When Mis12 and Ndc80 are combined and their structure is examined by negative stain or rotary shadowing EM, they appear as ~90-nm particles, indicating that they interact ‘in series’ [238,239] (Figure 4D). Recent crystal structures of the human and yeast complexes demonstrate that the four subunits of the Mis12 complex are structural paralogs with high helical content (Figure 4E).…”

Section: The Outer Kinetochorementioning

confidence: 95%

“…In the first mechanism, CENP-C directly binds to the Mis12 complex [165,238,240,241,245,278,279], which in turn binds to the Ndc80 complex and Knl1. In the second mechanism, the RWD domains in the Spc24 and Spc25 subunits of the Ndc80 complex directly interact with the intrinsically disordered N-terminal extension of CENP-T [75,113,239,242,279,280,281,282]. We summarize below the detailed understanding of these two mechanisms in yeast and vertebrates and their relative importance in outer kinetochore assembly in different systems.…”

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

“…The RWD domains in the Spc24 and Spc25 subunits of the Ndc80 complex interact directly with two related, short sequence motifs in the first 100 residues of the intrinsically disordered N-terminal extension of CENP-T (consisting of approximately 450 residues in humans) [75,113,239,242,279,280,281,282]. In humans, this interaction requires CDK phosphorylation of the CENP-T motifs, but the equivalent interaction of Spc24:Spc25 with CENP-T Cnn1 in S. cerevisiae may not require phosphorylation [242,279,281].…”

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

“…In humans, this interaction requires CDK phosphorylation of the CENP-T motifs, but the equivalent interaction of Spc24:Spc25 with CENP-T Cnn1 in S. cerevisiae may not require phosphorylation [242,279,281]. At least in vitro, the two Ndc80 complex-binding motifs of CENP-T can be occupied concomitantly, suggesting that this mechanism can recruit up to two Ndc80 complexes per CENP-T molecule [113,239]. The motifs on CENP-T are closely related to the Spc24:Spc25-binding motif in Dsn1 (discussed in the previous section) [242].…”

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

“…In an interesting recent twist, it was realized that CENP-T also contributes to kinetochore recruitment of the Mis12 complex [239,280,288]. This is promoted by a direct interaction of the Mis12 complex with a distinct, non-canonical CDK phosphorylation site on human CENP-T, Ser201 [239].…”

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

See 4 more Smart Citations

A Molecular View of Kinetochore Assembly and Function

Musacchio

Desai

2017

Biology

Self Cite

498

589

View full text Add to dashboard Cite

Kinetochores are large protein assemblies that connect chromosomes to microtubules of the mitotic and meiotic spindles in order to distribute the replicated genome from a mother cell to its daughters. Kinetochores also control feedback mechanisms responsible for the correction of incorrect microtubule attachments, and for the coordination of chromosome attachment with cell cycle progression. Finally, kinetochores contribute to their own preservation, across generations, at the specific chromosomal loci devoted to host them, the centromeres. They achieve this in most species by exploiting an epigenetic, DNA-sequence-independent mechanism; notable exceptions are budding yeasts where a specific sequence is associated with centromere function. In the last 15 years, extensive progress in the elucidation of the composition of the kinetochore and the identification of various physical and functional modules within its substructure has led to a much deeper molecular understanding of kinetochore organization and the origins of its functional output. Here, we provide a broad summary of this progress, focusing primarily on kinetochores of humans and budding yeast, while highlighting work from other models, and present important unresolved questions for future studies.

show abstract

Section: The Outer Kinetochorementioning

confidence: 95%

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

Section: Linkages Between the Inner And The Outer Kinetochorementioning

confidence: 99%

See 3 more Smart Citations

A Molecular View of Kinetochore Assembly and Function

Musacchio

Desai

2017

Biology

Self Cite

498

589

View full text Add to dashboard Cite

show abstract

Tension sensors reveal how the kinetochore shares its load

Salmon

Bloom

2017

BioEssays

View full text Add to dashboard Cite

At metaphase in mitotic cells, pulling forces at the kinetochore-microtubule interface create tension by stretching the centromeric chromatin between oppositely oriented sister kinetochores. This tension is important for stabilizing the end-on kinetochore microtubule attachment required for proper bi-orientation of sister chromosomes as well as for satisfaction of the Spindle Assembly Checkpoint and entry into anaphase. How force is coupled by proteins to kinetochore microtubules and resisted by centromere stretch is becoming better understood as many of the proteins involved have been identified. Recent application of genetically encoded fluorescent tension sensors within the mechanical linkage between the centromere and kinetochore microtubules are beginning to reveal – from live cell assays – protein specific contributions that are functionally important.

show abstract

Reconstitution and use of highly active human CDK1:Cyclin‐B:CKS1 complexes

et al. 2021

Self Cite

View full text Add to dashboard Cite

As dividing cells transition into mitosis, hundreds of proteins are phosphorylated by a complex of cyclin‐dependent kinase 1 (CDK1) and Cyclin‐B, often at multiple sites. CDK1:Cyclin‐B phosphorylation patterns alter conformations, interaction partners, and enzymatic activities of target proteins and need to be recapitulated in vitro for the structural and functional characterization of the mitotic protein machinery. This requires a pure and active recombinant kinase complex. The kinase activity of CDK1 critically depends on the phosphorylation of a Threonine residue in its activation loop by a CDK1‐activating kinase (CAK). We developed protocols to activate CDK1:Cyclin‐B either in vitro with purified CAKs or in insect cells through CDK‐CAK co‐expression. To boost kinase processivity, we reconstituted a ternary complex consisting of CDK1, Cyclin‐B, and CKS1. In this work, we provide and compare detailed protocols to obtain and use highly active CDK1:Cyclin‐B (CC) and CDK1:Cyclin‐B:CKS1 (CCC).

show abstract

Molecular basis of outer kinetochore assembly on CENP-T

Cited by 124 publications

References 61 publications

A Molecular View of Kinetochore Assembly and Function

A Molecular View of Kinetochore Assembly and Function

Tension sensors reveal how the kinetochore shares its load

Reconstitution and use of highly active human CDK1:Cyclin‐B:CKS1 complexes

Contact Info

Product

Resources

About