Phosphorylation within Intrinsic Disordered Region
Discriminates Histone Variant macroH2A1 Splicing
Isoforms-macroH2A1.1 and macroH2A1.2

J 2021

Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2

GIALLONGO, Sebastiano, Oriana LO RE, Gabriela LOCHMANOVÁ, Luca PARCA, Francesco PETRIZZELLI et. al.

Basic information

Original name

Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2

Authors

GIALLONGO, Sebastiano (380 Italy, belonging to the institution), Oriana LO RE (203 Czech Republic), Gabriela LOCHMANOVÁ (203 Czech Republic, belonging to the institution), Luca PARCA (380 Italy), Francesco PETRIZZELLI (380 Italy), Zbyněk ZDRÁHAL (203 Czech Republic, guarantor, belonging to the institution), Tommaso MAZZA (380 Italy) and Vinciguerra MANLIO (203 Czech Republic)

Edition

Biology, Basel, MDPI, 2021, 2079-7737

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30500 3.5 Other medical sciences

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

fulltext

Impact factor

Impact factor: 5.168

RIV identification code

RIV/00216224:14110/21:00119597

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.3390/biology10070659

UT WoS

000675987800001

Keywords in English

mass spectrometry; post-translational modifications

Abstract

V originále

Simple Summary MacroH2A1, a histone H2A variant, is present as two alternative splicing isoforms, macroH2A1.1 and macroH2A1.2, which are finely regulated through several mechanisms, including post-translational modifications (PTM). In this article, the authors provide the PTM pattern of macroH2A1.1 and macroH2A1.2 in the same experimental setting through mass spec analysis. They report a different phosphorylation level in their intrinsically disordered linker region, which can be responsible for their different biological role, as computational analysis shows. Background: Gene expression in eukaryotic cells can be governed by histone variants, which replace replication-coupled histones, conferring unique chromatin properties. MacroH2A1 is a histone H2A variant containing a domain highly similar to H2A and a large non-histone (macro) domain. MacroH2A1, in turn, is present in two alternatively exon-spliced isoforms: macroH2A1.1 and macroH2A1.2, which regulate cell plasticity and proliferation in a remarkably distinct manner. The N-terminal and the C-terminal tails of H2A histones stem from the nucleosome core structure and can be target sites for several post-translational modifications (PTMs). MacroH2A1.1 and macroH2A1.2 isoforms differ only in a few amino acids and their ability to bind NAD-derived metabolites, a property allegedly conferring their different functions in vivo. Some of the modifications on the macroH2A1 variant have been identified, such as phosphorylation (T129, S138) and methylation (K18, K123, K239). However, no study to our knowledge has analyzed extensively, and in parallel, the PTM pattern of macroH2A1.1 and macroH2A1.2 in the same experimental setting, which could facilitate the understanding of their distinct biological functions in health and disease. Methods: We used a mass spectrometry-based approach to identify the sites for phosphorylation, acetylation, and methylation in green fluorescent protein (GFP)-tagged macroH2A1.1 and macroH2A1.2 expressed in human hepatoma cells. The impact of selected PTMs on macroH2A1.1 and macroH2A1.2 structure and function are demonstrated using computational analyses. Results: We identified K7 as a new acetylation site in both macroH2A1 isoforms. Quantitative comparison of histone marks between the two isoforms revealed significant differences in the levels of phosphorylated T129 and S170. Our computational analysis provided evidence that the phosphorylation status in the intrinsically disordered linker region in macroH2A1 isoforms might represent a key regulatory element contributing to their distinct biological responses. Conclusions: Taken together, our results report different PTMs on the two macroH2A1 splicing isoforms as responsible for their distinct features and distribution in the cell.

Links

GF19-29701L, research and development project

Name: Funkce HDAC1 v T-buněčných lymfomech

Investor: Czech Science Foundation, Partner Agency (Austria)

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

MUNI/A/1325/2020, interní kód MU

Name: Biomedicínské vědy

Investor: Masaryk University

90127, large research infrastructures

Name: CIISB II

Citovat

GIALLONGO, Sebastiano, Oriana LO RE, Gabriela LOCHMANOVÁ, Luca PARCA, Francesco PETRIZZELLI, Zbyněk ZDRÁHAL, Tommaso MAZZA and Vinciguerra MANLIO. Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2. Biology. Basel: MDPI, 2021, vol. 10, No 7, p. 1-13. ISSN 2079-7737. Available from: https://dx.doi.org/10.3390/biology10070659.

@article{1824680,
   author = {Giallongo, Sebastiano and Lo Re, Oriana and Lochmanová, Gabriela and Parca, Luca and Petrizzelli, Francesco and Zdráhal, Zbyněk and Mazza, Tommaso and Manlio, Vinciguerra},
   article_location = {Basel},
   article_number = {7},
   doi = {http://dx.doi.org/10.3390/biology10070659},
   keywords = {mass spectrometry; post-translational modifications},
   language = {eng},
   issn = {2079-7737},
   journal = {Biology},
   title = {Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2},
   url = {https://www.mdpi.com/2079-7737/10/7/659},
   volume = {10},
   year = {2021}
}

TY  - JOUR
ID  - 1824680
AU  - Giallongo, Sebastiano - Lo Re, Oriana - Lochmanová, Gabriela - Parca, Luca - Petrizzelli, Francesco - Zdráhal, Zbyněk - Mazza, Tommaso - Manlio, Vinciguerra
PY  - 2021
TI  - Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2
JF  - Biology
VL  - 10
IS  - 7
SP  - 1-13
EP  - 1-13
PB  - MDPI
SN  - 20797737
KW  - mass spectrometry
KW  - post-translational modifications
UR  - https://www.mdpi.com/2079-7737/10/7/659
N2  - Simple Summary MacroH2A1, a histone H2A variant, is present as two alternative splicing isoforms, macroH2A1.1 and macroH2A1.2, which are finely regulated through several mechanisms, including post-translational modifications (PTM). In this article, the authors provide the PTM pattern of macroH2A1.1 and macroH2A1.2 in the same experimental setting through mass spec analysis. They report a different phosphorylation level in their intrinsically disordered linker region, which can be responsible for their different biological role, as computational analysis shows. Background: Gene expression in eukaryotic cells can be governed by histone variants, which replace replication-coupled histones, conferring unique chromatin properties. MacroH2A1 is a histone H2A variant containing a domain highly similar to H2A and a large non-histone (macro) domain. MacroH2A1, in turn, is present in two alternatively exon-spliced isoforms: macroH2A1.1 and macroH2A1.2, which regulate cell plasticity and proliferation in a remarkably distinct manner. The N-terminal and the C-terminal tails of H2A histones stem from the nucleosome core structure and can be target sites for several post-translational modifications (PTMs). MacroH2A1.1 and macroH2A1.2 isoforms differ only in a few amino acids and their ability to bind NAD-derived metabolites, a property allegedly conferring their different functions in vivo. Some of the modifications on the macroH2A1 variant have been identified, such as phosphorylation (T129, S138) and methylation (K18, K123, K239). However, no study to our knowledge has analyzed extensively, and in parallel, the PTM pattern of macroH2A1.1 and macroH2A1.2 in the same experimental setting, which could facilitate the understanding of their distinct biological functions in health and disease. Methods: We used a mass spectrometry-based approach to identify the sites for phosphorylation, acetylation, and methylation in green fluorescent protein (GFP)-tagged macroH2A1.1 and macroH2A1.2 expressed in human hepatoma cells. The impact of selected PTMs on macroH2A1.1 and macroH2A1.2 structure and function are demonstrated using computational analyses. Results: We identified K7 as a new acetylation site in both macroH2A1 isoforms. Quantitative comparison of histone marks between the two isoforms revealed significant differences in the levels of phosphorylated T129 and S170. Our computational analysis provided evidence that the phosphorylation status in the intrinsically disordered linker region in macroH2A1 isoforms might represent a key regulatory element contributing to their distinct biological responses. Conclusions: Taken together, our results report different PTMs on the two macroH2A1 splicing isoforms as responsible for their distinct features and distribution in the cell.
ER  -

GIALLONGO, Sebastiano, Oriana LO RE, Gabriela LOCHMANOVÁ, Luca PARCA, Francesco PETRIZZELLI, Zbyněk ZDRÁHAL, Tommaso MAZZA and Vinciguerra MANLIO. Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms-macroH2A1.1 and macroH2A1.2. \textit{Biology}. Basel: MDPI, 2021, vol.~10, No~7, p.~1-13. ISSN~2079-7737. Available from: https://dx.doi.org/10.3390/biology10070659.

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