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Gamuyao, R.

Publications and source records attributed to Gamuyao, R..

2 recordsLinked to original sources

Stress triggers global histone synthesis in the liver

Despite extensive drug development targeting DNA synthesis, there have been few efforts to inhibit histone synthesis, which occurs jointly with DNA synthesis in S-phase. A major roadblock is the lack of tools to quantitatively measure histone synthesis due to the high abundance and stability of histone proteins. Here we present chemical labeling methodologies to produce the first in vivo measurements of histone synthesis rates. Under basal conditions, histone synthesis tracks closely with tissue proliferation. However, multiple systemic and liver-specific stresses induce a previously uncharacterized surge of hepatocyte nucleosome production that is uncoupled from DNA replication. Tracing of newly synthesized nucleosomes detected promoter deposition, which was required to buffer transcriptional output. These results may inform strategies to modulate histone production to influence stress responses or disease.

molecular biology↗

A fluorogenic complementation tool kit for interrogating lipid droplet-organelle interaction

Contact sites between lipid droplets and other organelles are essential for cellular lipid and energy homeostasis. Detection of these contact sites at nanometer scale over time in living cells is challenging. Here, we developed a tool kit for detecting contact sites based on Fluorogen- Activated Bimolecular complementation at CONtact sites, FABCON, using a reversible, low affinity split fluorescent protein, splitFAST. FABCON labels contact sites with minimal perturbation to organelle interaction. Via FABCON, we quantitatively demonstrated that endoplasmic reticulum (ER)- and mitochondria (mito)-lipid droplet contact sites are dynamic foci in distinct metabolic conditions, such as during lipid droplet biogenesis and consumption. An automated analysis pipeline further classified individual contact sites into distinct subgroups based on size, likely reflecting differential regulation and function. Moreover, FABCON is generalizable to visualize a repertoire of organelle contact sites including ER-mito. Altogether, FABCON reveals insights into the dynamic regulation of lipid droplet-organelle contact sites and generates new hypotheses for further mechanistical interrogation during metabolic switch.

cell biology↗