Cell Lysate Microarray for Mapping the Network of Genetic Regulators for Histone Marks

Protein, as the major executer for cell progresses and functions, its abundance and the level of post-translational modifications, are tightly monitored by regulators. Genetic perturbation could help us to understand the relationships between genes and protein functions. Herein, we developed a cell lysate microarray on kilo-conditions (CLICK) from 4,837 yeast knockout (YKO) strains and 322 temperature-sensitive mutant strains to explore the impact of the genome-wide interruption on certain protein. Taking histone marks as examples, a general workflow was established for the global identification of upstream regulators. Through a single CLICK array test, we obtained a series of regulators for H3K4me3 which covers most of the known regulators in Saccharomyces cerevisiae. We also noted that several group of proteins that are linked to negatively regulation of H3K4me3. Further, we discovered that Cab4p and Cab5p, two key enzymes of CoA biosynthesis, play central roles in histone acylation. Because of its general applicability, CLICK array could be easily adopted to rapid and global identification of upstream protein/enzyme(s) that regulate/modify the level of a protein or the posttranslational modification of a non-histone protein.