# Dissecting mechanisms of transcriptional regulation during stress

> **NIH NIH R35** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2024 · $374,979

## Abstract

PROJECT SUMMARY
Mammalian cells are constantly challenged by a multitude of environmental stresses. In response, cells trigger
corrective measures to make critical cellular decisions. Dysregulation of these stress response programs are
associated with ageing, neurodegeneration, and cancer. However, the molecular basis of cellular stress
response is largely unknown. While stressed cells globally suppress RNA and protein synthesis, they
specifically induce stress response genes that determine cell fate. The overarching goal of my research is to
understand how mammalian cells regulate gene expression during stress. In this proposed work, we will
investigate the mechanism of two conserved and ill-understood gene regulatory features of stress response –
(1) accumulation of gene regulatory biomolecules within membraneless compartments, called condensates, and
(2) widespread disruption of transcription termination. We recently discovered that cells rapidly accumulate their
multivalent proteome, especially transcription modulators, within subcellular condensates during stress. While
the functions of condensates remain nebulous, it’s been shown that dysregulation or aggregation of condensate
constituents are associated with neurodegeneration. Building on our discovery, we found that heat shock factors
(HSFs), multivalent transcription factors and essential arbiters of proteostasis, form stress-induced nuclear
condensates that localize at cellular sites of elevated transcription. Our data suggest that stress-induced HSF
condensates intensely activate transcription at specific genomic loci and act as transcription hubs. We will
employ this exemplary system to gain a broader understanding of how stress-induced transcriptional
condensates function and how spatial organization drives transcriptional control. Additionally, we found that
disruption of transcription initiation and termination are correlated with sequestration of RNA metabolism
regulators and linked to global transcriptional downregulation during stress. Yet, disrupted termination at specific
genomic loci leads to extensive transcriptional readthrough via unclear mechanisms, culminating in the synthesis
of novel stress-specific transcripts. This phenomenon is also a hallmark of viral infections and renal cancers.
Together, these findings lead to the following questions – (1) How do stress-induced HSF condensates form
and function? We will employ a combination of advanced imaging and spatial-omics technologies that identify
molecular drivers of condensate formation and probe condensate’s function in transcriptional regulation within
cellular models of proteotoxic stress. (2) How do cells mediate stress-associated transcription
readthrough? Using nascent RNA sequencing and epigenomics we will identify epigenetic drivers of
readthrough in validated cellular models of osmotic and proteotoxic stress. We will then use single-molecule
imaging to elucidate the molecular drivers of transcriptional readthro...

## Key facts

- **NIH application ID:** 10941800
- **Project number:** 1R35GM155432-01
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Sethuramasundaram Pitchiaya
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $374,979
- **Award type:** 1
- **Project period:** 2024-06-25 → 2029-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10941800

## Citation

> US National Institutes of Health, RePORTER application 10941800, Dissecting mechanisms of transcriptional regulation during stress (1R35GM155432-01). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/10941800. Licensed CC0.

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