The cryo-EM structure of the CENP-A nucleosome in complex with ggKNL2

Jiang, Honghui; Ariyoshi, Mariko; Watanabe, Reito; Makino, Fumiaki; Namba, Keiichi; Fukagawa, Tatsuo

doi:10.1101/2022.06.24.497480

Cited by 2 publications

(6 citation statements)

References 69 publications

(133 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, we created a chimeric model combining AlphaFold2 predictions, which was independently validated twice with molecular dynamics for the stability (methods). This model aligns with the experimentally resolved structures, like the αKNL2 CENPC-k motif binds to histones similar to ggKNL2 (Figure 5A) (Jiang, et al 2022). Both SANTA domains interact with each other and contact all histones and DNA.…”

Section: βKnl2 Is Highly Dynamic and Loosely Bound To Centromeric Nuc...supporting

confidence: 85%

“…Previous studies have shown that the M18BP1/KNL2 requires the CENPC-like motif for centromeric localization (Hori, et al 2017;Sandmann, et al 2017;Smith and Maddox 2017;Jiang, et al 2022).…”

Section: βKnl2 Depends On Cenpc-like Motif Containing Knl2 For Centr...mentioning

confidence: 98%

“…Some previously predicted disordered regions of the histones and of αKNL2 were neglected in the prediction of the complex to decrease computational costs (Supplementary Figure S4). We superposed this prediction with the chicken CENP-A nucleosome in complex with KNL2, recently resolved by cryo-EM (pdb code 7y71) (Jiang, et al 2022) . AlphaFold2 correctly placed the CENPC-k motif of αKNL2 in a similar position of the experimentally resolved CENPC-like motif of chicken KNL2 (Supplementary Figure S3).…”

Section: Alphafold2 Protein Predictions and Molecular Dynamics Analysismentioning

confidence: 99%

“…Different inner kinetochore proteins associate with the centromeric DNA (Cheeseman 2014;Yamagishi, et al 2014;Musacchio and Desai 2017;Jiang, et al 2022) . For example, αKNL2 associates with DNA in a sequence-independent manner in vitro and preferentially with pAL1 in vivo (Sandmann, et al 2017;Yalagapati, et al 2024).…”

Section: βKnl2 Binds To the Centromeric Dna In Vitromentioning

confidence: 99%

“…The SANTA of Xenopus M18BP1/KNL2 mediates its interaction with CENP-C during the metaphase, thus regulating its centromere targeting (French and Straight 2019). However, M18BP1/KNL2 interaction with CENP-A/CENH3 nucleosomes in Xenopus and chicken does not require the SANTA (French, et al 2017;Jiang, et al 2022). Whereas modification of the CENPC-like motif in Arabidopsis, chicken and Xenopus led to mis-localization of KNL2 (French, et al 2017;Hori, et al 2017;Sandmann, et al 2017).…”

Section: Introductionmentioning

confidence: 98%

See 4 more Smart Citations

Structural Basis of βKNL2 Centromeric Targeting Mechanism and Its Role in Plant-Specific Kinetochore Assembly

Yadala,

Camara,

Yalagapati

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

The kinetochore is an essential protein complex that ensures proper chromosome segregation during cell division. Kinetochore assembly is initiated by the incorporation of CENP-A/CENH3. This process depends on KNL2/M18BP1 and CENP-C proteins. In plants, two variants of KNL2, αKNL2 and βKNL2, are present. Both possess the conserved SANTA domain, while αKNL2 additionally has the centromere-targeting CENPC-k motif. Despite lacking the CENPC-like motif, the plant-specific βKNL2 localizes to centromeres and aids in CENP-A/CENH3 loading. We found that efficient centromeric targeting of βKNL2 requires the SANTA domain and the C-terminal part, while nuclear targeting depends on a conserved C-terminal motif-III. Structural predictions and experimental validations reveal that βKNL2 forms homodimers and interacts with centromeric DNA and αKNL2. We confirm that centromeric targeting of βKNL2 depends on αKNL2 in a tissue-dependent manner. Our findings provide crucial insights into the unique mechanisms of plant-specific kinetochore assembly, highlighting βKNL2’s essential role in this process.Graphical abstract

show abstract

Section: βKnl2 Is Highly Dynamic and Loosely Bound To Centromeric Nuc...supporting

confidence: 85%

Section: βKnl2 Depends On Cenpc-like Motif Containing Knl2 For Centr...mentioning

confidence: 98%

Section: Alphafold2 Protein Predictions and Molecular Dynamics Analysismentioning

confidence: 99%

Section: βKnl2 Binds To the Centromeric Dna In Vitromentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 3 more Smart Citations

Structural Basis of βKNL2 Centromeric Targeting Mechanism and Its Role in Plant-Specific Kinetochore Assembly

Yadala,

Camara,

Yalagapati

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Direct recruitment of Mis18 to interphase spindle poles promotes CENP-A chromatin assembly

London,

Medina-Pritchard,

Spanos

et al. 2023

Preprint

View full text Add to dashboard Cite

CENP-A chromatin specifies mammalian centromere identity, and its chaperone HJURP replenishes CENP-A when recruited by the Mis18 complex (Mis18C) via M18BP/KNL2 to CENP-C at kinetochores during interphase. However, the Mis18C recruitment mechanism remains unresolved in species lacking M18BP1, such as fission yeast. Fission yeast centromeres cluster at G2 spindle pole bodies (SPBs) when CENP-ACnp1 is replenished and where Mis18C also localizes. We show that SPBs play an unexpected role in concentrating Mis18C near centromeres through the recruitment of Mis18 by direct binding to the major SPB LInker of Nucleoskeleton and Cytoskeleton (LINC) complex component Sad1. Mis18 recruitment by Sad1 is important for CENP-ACnp1 chromatin establishment and acts in parallel with a CENP-C-mediated Mis18C recruitment pathway to maintain centromeric CENP-ACnp1, but is independent of Sad1-mediated centromere clustering. SPBs therefore provide a non-chromosomal scaffold for both Mis18C recruitment and centromere clustering during G2. This centromere-independent Mis18-SPB recruitment provides a mechanism that governs de novo CENP-ACnp1 chromatin assembly by the proximity of appropriate sequences to SPBs and highlights how nuclear spatial organization influences centromere identity.

show abstract

The cryo-EM structure of the CENP-A nucleosome in complex with ggKNL2

Cited by 2 publications

References 69 publications

Structural Basis of βKNL2 Centromeric Targeting Mechanism and Its Role in Plant-Specific Kinetochore Assembly

Structural Basis of βKNL2 Centromeric Targeting Mechanism and Its Role in Plant-Specific Kinetochore Assembly

Direct recruitment of Mis18 to interphase spindle poles promotes CENP-A chromatin assembly

Contact Info

Product

Resources

About