Format

Send to

Choose Destination
G3 (Bethesda). 2017 Dec 4;7(12):3857-3866. doi: 10.1534/g3.117.300252.

Dissecting Nucleosome Function with a Comprehensive Histone H2A and H2B Mutant Library.

Jiang S1,2,3, Liu Y1,2, Xu C1,2, Wang Y4,5, Gong J4,5, Shen Y4,5, Wu Q1,2, Boeke JD6,7, Dai J8,2,3.

Author information

1
Ministry of Education Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, PR China.
2
Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua University, Beijing 100084, PR China.
3
Center for Synthetic Biology Engineering Research, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
4
China National GeneBank, Beijing Genomics Institute-Shenzhen, Shenzhen 518120, China.
5
Beijing Genomics Institute-Shenzhen, Shenzhen 518083, China.
6
Institute for Systems Genetics, New York University Langone Medical Center, New York 10011.
7
Department of Biochemistry and Molecular Pharmacology, New York University Langone Medical Center, New York 10011.
8
Ministry of Education Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, PR China junbiao.dai@siat.ac.cn.

Abstract

Using a comprehensive library of histone H2A and H2B mutants, we assessed the biological function of each amino acid residue involved in various stress conditions including exposure to different DNA damage-inducing reagents, different growth temperatures, and other chemicals. H2B N- and H2A C-termini were critical for maintaining nucleosome function and mutations in these regions led to pleiotropic phenotypes. Additionally, two screens were performed using this library, monitoring heterochromatin gene silencing and genome stability, to identify residues that could compromise normal function when mutated. Many distinctive regions within the nucleosome were revealed. Furthermore, we used the barcode sequencing (bar-seq) method to profile the mutant composition of many libraries in one high-throughput sequencing experiment, greatly reducing the labor and increasing the capacity. This study not only demonstrates the applications of the versatile histone library, but also reveals many previously unknown functions of histone H2A and H2B.

KEYWORDS:

DNA damage; heterochromatin gene silencing; histone; post-translational modification

PMID:
29038170
PMCID:
PMC5714483
DOI:
10.1534/g3.117.300252
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center