Deciphering histone codes underlying diurnal and cell cycle state transitions in Chlamydomonas
EMSL Project ID
51056
Abstract
The long-term goal of this project is to understand the dynamic genetic programming which underlies cell growth and proliferation in green algae. Proliferating microalgae in a diurnal environment are in a constant state of flux as they undergo changes in light-cued and dark-cued gene expression and metabolic states in coordination with cell division. One of the major challenges for systems biology and predictive modeling is our incomplete understanding of gene expression networks and underlying chromatin dynamics, particularly for green algae which have lineage-specific features that cannot be extrapolated from work done in animals, fungi or even land plants. We propose here to leverage the powerful features of Chlamydomonas reinhardtii (Chlamydomonas) as a well-developed model for cell cycle regulation in the green lineage, our expertise in synchronizing and controlling the cell cycle of Chlamydomonas, expertise in Chlamydomonas histones, and the unique expertise and instrumentation of EMSL in analysis of histone post-translational modification (PTMs) and intact protein complexes to gain insight into the life cycle dynamics of Chlamydomonas histones and chromatin. This project also addresses a related question and long-standing enigma of how the Chlamydomonas retinoblastoma tumor suppressor complex (RBC), a conserved chromatin-associated cell cycle regulator, is modulated during the cell cycle and how it impacts chromatin dynamics. Under this collaborative proposal we will use EMSL's state-of-the-art instrumentation and expertise to: i) profile histone PTMs across the synchronous cell cycle of Chlamydomonas; ii) determine how global histone PTMs are impacted in cell cycle mutants that are missing RBC subunits; iii) determine the composition and PTMs of purified RBC from different cell cycle stages to assess how it is modified during the cell cycle, and determine whether RBC-associated histones are enriched for specific PTMs. Data from this proposal will provide a ground breaking and high-quality foundational resource which will quantitatively describe the unique set of histone marks and their dynamic changes during the Chlamydomonas life cycle. In conjunction, we will begin to gain new insight into the regulation and cell cycle control mechanism of the RBC that will help resolve long-standing questions about how it impacts chromatin and effects cell cycle state changes.
Project Details
Project type
Exploratory Research
Start Date
2019-11-26
End Date
2021-03-31
Status
Closed
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
Team Members
Related Publications
Bradley S. Evans, Ljiljana Paša-Tolić, James J. Pesavento, Sarah R. Rommelfanger, Henna Shaghasi, Shin-Cheng Tzeng, James G. Umen, Mowei Zhou. 2021. "An Improved Top-Down Mass Spectrometry Characterization of Chlamydomonas reinhardtii Histones and Their Post-translational Modifications." Journal of the American Society for Mass Spectrometry 32 (7):1671-1688. https://doi.org/10.1021/jasms.1c00029