Stability of nucleosomes containing homogenously ubiquitylated H2A and H2B prepared using semisynthesis

Beat Fierz; Sinan Kilic; Aaron R. Hieb; Karolin Luger; Tom W. Muir

doi:10.1021/ja308908p

Stability of nucleosomes containing homogenously ubiquitylated H2A and H2B prepared using semisynthesis

Beat Fierz, Sinan Kilic, Aaron R. Hieb, Karolin Luger, Tom W. Muir

Research output: Contribution to journal › Article › peer-review

82 Scopus citations

Abstract

Post-translational modifications (PTMs) of histones are an essential feature in the dynamic regulation of chromatin. One of these modifications, ubiquitylation, has been speculated to directly influence the stability of the nucleosome, which represents the basic building block of chromatin. Here we report a strategy for the semisynthesis of site-specifically ubiquitylated histone H2A (uH2A). This branched protein was generated through a three-piece expressed protein ligation approach including a traceless ligation at valine. uH2A could be efficiently incorporated into nucleosomes, thereby opening the way to detailed biochemical and biophysical studies on the function of this PTM. Accordingly, we used uH2A, as well as a previously generated ubiquitylated H2B, in chaperone-coupled nucleosome stability assays to demonstrate that the direct effect of ubiquitylated histones on nucleosomal stability is in fact modest.

Original language	English (US)
Pages (from-to)	19548-19551
Number of pages	4
Journal	Journal of the American Chemical Society
Volume	134
Issue number	48
DOIs	https://doi.org/10.1021/ja308908p
State	Published - Dec 5 2012

All Science Journal Classification (ASJC) codes

General Chemistry
Biochemistry
Catalysis
Colloid and Surface Chemistry

Access to Document

10.1021/ja308908p

Cite this

@article{1dc10fb7f29e4dbf9cd93a0b90e671e6,

title = "Stability of nucleosomes containing homogenously ubiquitylated H2A and H2B prepared using semisynthesis",

abstract = "Post-translational modifications (PTMs) of histones are an essential feature in the dynamic regulation of chromatin. One of these modifications, ubiquitylation, has been speculated to directly influence the stability of the nucleosome, which represents the basic building block of chromatin. Here we report a strategy for the semisynthesis of site-specifically ubiquitylated histone H2A (uH2A). This branched protein was generated through a three-piece expressed protein ligation approach including a traceless ligation at valine. uH2A could be efficiently incorporated into nucleosomes, thereby opening the way to detailed biochemical and biophysical studies on the function of this PTM. Accordingly, we used uH2A, as well as a previously generated ubiquitylated H2B, in chaperone-coupled nucleosome stability assays to demonstrate that the direct effect of ubiquitylated histones on nucleosomal stability is in fact modest.",

author = "Beat Fierz and Sinan Kilic and Hieb, {Aaron R.} and Karolin Luger and Muir, {Tom W.}",

year = "2012",

month = dec,

day = "5",

doi = "10.1021/ja308908p",

language = "English (US)",

volume = "134",

pages = "19548--19551",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society",

number = "48",

}

TY - JOUR

T1 - Stability of nucleosomes containing homogenously ubiquitylated H2A and H2B prepared using semisynthesis

AU - Fierz, Beat

AU - Kilic, Sinan

AU - Hieb, Aaron R.

AU - Luger, Karolin

AU - Muir, Tom W.

PY - 2012/12/5

Y1 - 2012/12/5

N2 - Post-translational modifications (PTMs) of histones are an essential feature in the dynamic regulation of chromatin. One of these modifications, ubiquitylation, has been speculated to directly influence the stability of the nucleosome, which represents the basic building block of chromatin. Here we report a strategy for the semisynthesis of site-specifically ubiquitylated histone H2A (uH2A). This branched protein was generated through a three-piece expressed protein ligation approach including a traceless ligation at valine. uH2A could be efficiently incorporated into nucleosomes, thereby opening the way to detailed biochemical and biophysical studies on the function of this PTM. Accordingly, we used uH2A, as well as a previously generated ubiquitylated H2B, in chaperone-coupled nucleosome stability assays to demonstrate that the direct effect of ubiquitylated histones on nucleosomal stability is in fact modest.

AB - Post-translational modifications (PTMs) of histones are an essential feature in the dynamic regulation of chromatin. One of these modifications, ubiquitylation, has been speculated to directly influence the stability of the nucleosome, which represents the basic building block of chromatin. Here we report a strategy for the semisynthesis of site-specifically ubiquitylated histone H2A (uH2A). This branched protein was generated through a three-piece expressed protein ligation approach including a traceless ligation at valine. uH2A could be efficiently incorporated into nucleosomes, thereby opening the way to detailed biochemical and biophysical studies on the function of this PTM. Accordingly, we used uH2A, as well as a previously generated ubiquitylated H2B, in chaperone-coupled nucleosome stability assays to demonstrate that the direct effect of ubiquitylated histones on nucleosomal stability is in fact modest.

UR - http://www.scopus.com/inward/record.url?scp=84870708220&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84870708220&partnerID=8YFLogxK

U2 - 10.1021/ja308908p

DO - 10.1021/ja308908p

M3 - Article

C2 - 23163596

AN - SCOPUS:84870708220

SN - 0002-7863

VL - 134

SP - 19548

EP - 19551

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 48

ER -

Stability of nucleosomes containing homogenously ubiquitylated H2A and H2B prepared using semisynthesis

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this