# Structural and functional determinants of biomolecular condensates in transcription organization

> **NIH NIH R35** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2024 · $388,043

## Abstract

Many cellular processes use phase separation to sequesters biomolecules into membraneless organelles
or condensates. This concept is of particular importance in the cell nucleus. Most nuclear condensates
are associated with specific chromatin loci. Thus, chromatin organization and biomolecular condensate
formation are closely related. Topology, epigenetic modification, or enzymatic activity are possible links
between the chromatin scaffold and protein condensation. We have previously described condensates
that concentrate transcription machinery at super-enhancers but understanding of cell-to-cell variability
and locus specific features that nucleate condensates is missing. Preliminary data indicates that active
chromatin decorates the surface of transcription condensates below the resolution limit of conventional
microscopy. Making use of recent developments in multiplexed super-resolution microscopy of both
chromatin elements and protein factors, we will characterize the nature of the chromatin enrichment
layer and determine if it reflects the condensate surface as an active interface that regulates
transcription. We will for the first time directly observe the interplay of local chromatin topology,
epigenetic signatures, and condensate formation in a specific model locus in mouse embryonic stem
cells. By scrutinizing with genomic resolution how each of the regulatory elements in the locus interacts
with the condensate we will determine if chromatin structures such as enhancer hubs nucleate
condensates, or if condensates induce specific chromatin topologies by connecting active elements.
Finally, we will investigate at the molecular level how condensate constituents interact with the
microenvironment and associated chromatin elements, and test the hypothesis that retention of
transcription factors boosts binding site occupancy and stabilizes condensates at regulatory chromatin.
Targeted perturbation and locus-specific observation will allow us to answer this question and begin to
identify genome-wide rules that predict where precisely chromatin-supported condensates form.
The overall vision for this research program is to develop an integrated view of chromatin organization
and condensate formation. Experimental data will inspire new conceptual frameworks and help us
integrate the full complexity of condensates into paradigms of biological function such as transcription
regulation. We expect that new biophysical models of self-organization and self-regulation will emerge
from our studies of intracellular condensates.

## Key facts

- **NIH application ID:** 10933412
- **Project number:** 5R35GM150560-02
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** Jan-Hendrik Spille
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $388,043
- **Award type:** 5
- **Project period:** 2023-09-22 → 2028-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10933412

## Citation

> US National Institutes of Health, RePORTER application 10933412, Structural and functional determinants of biomolecular condensates in transcription organization (5R35GM150560-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10933412. Licensed CC0.

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