Native and Tagged CENP-A Histones Are Functionally Inequivalent

Overview

Journal Epigenetics Chromatin

Publisher Biomed Central

Specialties Biochemistry
Genetics

Date 2024 Jun 2

PMID 38825690

Authors

Minh Bui

Songjoon Baek

Reda S Bentahar

Daniel P Melters

Yamini Dalal

Affiliations

Soon will be listed here.

Abstract

Background: Over the past several decades, the use of biochemical and fluorescent tags has elucidated mechanistic and cytological processes that would otherwise be impossible. The challenging nature of certain nuclear proteins includes low abundancy, poor antibody recognition, and transient dynamics. One approach to get around those issues is the addition of a peptide or larger protein tag to the target protein to improve enrichment, purification, and visualization. However, many of these studies were done under the assumption that tagged proteins can fully recapitulate native protein function.

Results: We report that when C-terminally TAP-tagged CENP-A histone variant is introduced, it undergoes altered kinetochore protein binding, differs in post-translational modifications (PTMs), utilizes histone chaperones that differ from that of native CENP-A, and can partially displace native CENP-A in human cells. Additionally, these tagged CENP-A-containing nucleosomes have reduced centromeric incorporation at early G1 phase and poorly associates with linker histone H1.5 compared to native CENP-A nucleosomes.

Conclusions: These data suggest expressing tagged versions of histone variant CENP-A may result in unexpected utilization of non-native pathways, thereby altering the biological function of the histone variant.

Citing Articles

High-resolution analysis of human centromeric chromatin.

Melters D, Bui M, Rakshit T, Grigoryev S, Sturgill D, Dalal Y Life Sci Alliance. 2025; 8(4.

PMID: 39848706 PMC: 11757159. DOI: 10.26508/lsa.202402819.

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