# Investigating Circadian Post-Transcriptional Regulation using Widefield Microscopy.

> **NIH NIH R35** · RENSSELAER POLYTECHNIC INSTITUTE · 2024 · $119,971

## Abstract

Project Summary/Abstract:
 Circadian rhythms are highly conserved, 24-hour, oscillations that tune human physiology to the
day/night cycle, enhancing fitness by ensuring that appropriate activities occur at biologically advantageous
times. Disruption of proper circadian synchronization negatively impacts the human long-term medical outlook,
making revealing the mechanism underlying circadian regulation over cellular physiology critical to human
health. Circadian rhythms are timed via a transcription-translation based negative feedback loop (TTFL), or
clock. The current paradigm for circadian regulation over physiology, termed the clocks “output”, is that
transcriptional programing generated by the TTFL drives temporally specific waves of gene expression.
However, our research has revealed that transcriptional programing cannot wholly account for clock output, as
we discovered little to no correlation between mRNAs and proteins that oscillate with a circadian periodicity in
fungi and macrophages, revealing robust circadian post-transcriptional control over immunometabolism. While
the mechanisms that time this circadian post-transcriptional control are unknown, we have demonstrated the
proteins involved in the transcriptionally repressive arm of the clock act as hub proteins for the formation of
large macromolecular complexes. Many of the proteins bound to these complexes are circadianly controlled at
the post-transcriptional level. Moreover, we established the clock repressive arm proteins have conserved
regions of intrinsic protein disorder that correlate with the predicted binding sights of the identified interactors.
 Our research goal is to validate the hypothesis that circadian post-transcriptional regulation is tuned via
the formation of time-of-day defined macromolecular protein complexes centered around the repressive arm
proteins. We posit that the formation of these complexes is enabled by conserved intrinsic disorder in the
repressive arm proteins, allowing flexibility to widely impart circadian post-transcriptional regulation over
immunometabolism. To test our hypothesis, we will create a Conformational/Temporal Interactome (CiTI) map
of clock repressive complex proteins in mammals and fungi. We will generate these CiTI maps in a sex and
immune-specific manner to define the effect of post-transcriptional circadian control over physiology. As a
mechanism for keeping time, circadian feedback loops are highly conserved and much of the definition of clock
mechanisms comes from the investigation of model systems. We will therefore exploit cost-effective fungal and
mammalian model systems to address our hypotheses. This supplement will support the purchase of a widefield
microscope, which will help to address all of the proposed hypotheses above, furthering our long-term goal of
elucidating the fundamental principles of circadian timing, revealing the mechanisms of circadian control over
cellular physiology.

## Key facts

- **NIH application ID:** 11032133
- **Project number:** 3R35GM128687-06S1
- **Recipient organization:** RENSSELAER POLYTECHNIC INSTITUTE
- **Principal Investigator:** Jennifer Marie Hurley
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $119,971
- **Award type:** 3
- **Project period:** 2018-08-15 → 2028-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11032133

## Citation

> US National Institutes of Health, RePORTER application 11032133, Investigating Circadian Post-Transcriptional Regulation using Widefield Microscopy. (3R35GM128687-06S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/11032133. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*