Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Sep 9;8:90. doi: 10.1038/s41421-022-00439-6

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 2 — a The 3.7 Å resolution cryo-EM density map of CCAN–DNA complex at two different views. The density map of DNA is colored by hotpink and the others are shown in gray transparent surfaces. b Electrostatic potential surface view of CENP-LN-HIK^head-TW binding with DNA. The DNA is shown as cartoon. Note that positively charged amino acids from CENP-LN, CENP-I and CENP-TW constitute the contact sites between CCAN and DNA. c, d Comparison of elution profiles (c) of CCAN^ΔCT-CENP-A¹⁶⁷/CCAN^ΔCT-CENP-A^{167-(R80A-G81A)} and CCAN^6E-ΔCT-CENP-A¹⁶⁷ in Superose 6 5/150 GL (GE Health) and the Coomassie-blue stained 15% SDS-PAGE gel. CENP-A¹⁶⁷ is the CENP-A nucleosome reconstituted by using a DNA fragment of 167 bp in length. d The CCAN^ΔCT complex bound to either CENP-A nucleosome or CENP-A^R80A-G81A nucleosome which reconstituted with 167 bp DNA, but the CCAN^6E-ΔCT complex failed. The CCAN^ΔCT complex includes CENP-LN, CENP-HIKM and CENP-OPQUR, but not CENP-C and CENP-TWSX; the CCAN^6E-ΔCT complex includes charge mutations of positively-charged residues on the CENP-LN (K270E/K296E in CENP-N^2E, K155E/R306E/K319E/K321E in CENP-L^4E) in contact with DNA; two degradation products of CENP-R annotated as CENP-R^Δ1 and CENP-R^Δ2 in gray color. Of note, the two separated peaks seen in the elution represent wild type CCAN (red line) and CENP-A^Nuc–binding deficient CCAN (green line) complex with CENP-A^Nuc and CENP-A^Nuc which are indistinguishable. However, DNA binding-deficient CCAN (cyan line) failed to bind nucleosomal DNA, validating that CCAN binds to DNA via CENP-LN. See also Supplementary Figs. S5, S6.