# Rhythmic Dynamics of Stress Granules in Wild-Type and Bmal1−/− Fibroblasts Lacking a Functional Canonical Circadian Clock

## TL;DR

Stress granules show daily rhythms in mouse cells, even without a key circadian clock gene, suggesting alternative timing mechanisms.

## Contribution

Demonstrates that stress granule rhythmicity persists in Bmal1−/− cells, indicating non-canonical circadian regulation.

## Key findings

- SG number, eIF3 signal intensity, and area oscillated with ~24 h rhythms in NIH/3T3 cells.
- SG rhythmicity persisted in Bmal1−/− MEFs but with altered amplitude and phase.
- Several SG-associated RNA-binding proteins showed time-dependent changes in mRNA or protein levels.

## Abstract

Circadian rhythms are endogenous ~24 h oscillations that regulate diverse biochemical processes. Although stress responses can exhibit circadian modulation, evidence for rhythmic regulation of stress granules (SGs)—cytoplasmic RNA–protein condensates formed under stress—remains limited. We investigated sodium arsenite-induced SG dynamics in NIH/3T3 cultures. SG number, eIF3 signal intensity—an established SG marker—and area oscillated with a period of ~24 h. These rhythms persisted in Bmal1−/− mouse embryonic fibroblasts (MEFs), despite lacking a transcription–translation feedback loop (TTFL) that constitutes the canonical circadian clock, but with altered amplitude and phase, indicating partial dependence on the molecular clock. Several SG-associated RNA-binding proteins (TIA-1, BRF1, hnRNP Q, and LARK) exhibited time-dependent changes at the mRNA and/or protein level, suggesting potential mechanisms for rhythmic SG modulation. Unlike previous in vivo reports linking SG variation to eIF2α phosphorylation, no temporal changes in phosphorylated eIF2α were observed, highlighting differences between isolated cells and tissues. Our results show that SG rhythmicity can persist without BMAL1, supporting alternative oscillatory mechanisms that contribute to the temporal organization of stress responses. Given their role in cell survival and the association of SG dysfunction with disease, these rhythms provide insight into how cellular stress responses are temporally regulated.