Histone Methylomics and the Chromatin Landscape of Chlamydomonas reinhardtii
EMSL Project ID
60418
Abstract
Our goal is to identify and categorize the SET domain methyltransferase enzymes in the model unicellular green alga Chlamydomonas reinhardtii (Chlamydomonas) that are responsible for histone methylations--a group of post-translational modifications (PTMs) that play an important and critical role in eukaryotic chromatin dynamics and gene expression. The Chlamydomonas genome encodes 52 SET domain proteins, a family of lysine methyltransferases, some of which are responsible for methylation at different lysine residues within histone tails. Only two SET domain proteins have been biochemically characterized Chlamydomonas, and do not account for all of the known methylation marks on its histones (potentially up to 40 variants). Under this proposal we will comprehensively characterize Chlamydomonas SET domain proteins and identify the subset that have histone methyltransferase (HMT) activity, and begin to determine their contribution to chromatin structure and gene expression. Under Aim 1 we will use synthetic genes encoding Chlamydomonas SET domain proteins for in vivo localization studies to identify those that are nucleus-targeted, and for production of tagged recombinant proteins that will be purified and screened for histone methyltransferase activity in vitro. Under Aim 2 we will use either existing insertional mutants or create gene-edited mutants corresponding to positive candidates from Aim 1 to screen in vivo for histone PTM phenotypes, gene expression phenotypes, and chromatin phenotypes using a combination of mass spectrometry, transcriptomics and chromatin structural profiling. This project synergistically employs expertise from EMSL and JGI and will provide a key resource for understanding the language of algal chromatin and the impact of specific methylation marks on gene expression and other chromatin-related phenotypes.
Project Details
Project type
FICUS Research
Start Date
2022-10-01
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator
Co-Investigator(s)