Title

Characterization of Chlamydomonas reinhardtii Core Histones by Top-Down Mass Spectrometry Reveals Unique Algae-Specific Variants and Post-Translational Modifications

SMC Author

James Pesavento

SMC Affiliated Work

1

Status

Faculty

School

School of Science

Department

Biology

Document Type

Article

Publication Date

2017

Publication Title

Journal of Proteome Research

Description/Abstract

The unicellular microalga Chlamydomonas reinhardtii has played an instrumental role in the development of many new fields (bioproducts, biofuels, etc.) as well as the advancement of basic science (photosynthetic apparati, flagellar function, etc.). Chlamydomonas' versatility ultimately derives from the genes encoded in its genome and the way that the expression of these genes is regulated, which is largely influenced by a family of DNA binding proteins called histones. We characterize C. reinhardtii core histones, both variants and their post-translational modifications, by chromatographic separation, followed by top-down mass spectrometry (TDMS). Because TDMS has not been previously used to study Chlamydomonas proteins, we show rampant artifactual protein oxidation using established nuclei purification and histone extraction methods. After addressing oxidation, both histones H3 and H4 are found to each have a single polypeptide sequence that is minimally acetylated and methylated. Surprisingly, we uncover a novel monomethylation at lysine 79 on histone H4 present on all observed molecules. Histone H2B and H2A are found to have two and three variants, respectively, and both are minimally modified. This study provides an updated assessment of the core histone proteins in the green alga C. reinhardtii by top-down mass spectrometry and lays the foundation for further investigation of these essential proteins.

Keywords

epigenetics, histones, microalgae, post-translational modifications, top-down mass spectrometry

Scholarly

yes

DOI

10.1021/acs.jproteome.7b00780

Disciplines

Biology

Original Citation

Khan, A, Eikani, C. K., Khan, H., Iavarone, A. T., and Pesavento, J. J. “Characterization of Chlamydomonas reinhardtii core histones by top-down mass spectrometry reveals unique algae-specific variants and post-translational modifications.” Journal of Proteome Research. 2017.

Share

COinS