# The Role of G Protein-coupled Receptors in Red Tide Dinoflagellate Bioluminescence

> **NIH NIH R21** · UNIVERSITY OF SOUTH FLORIDA · 2023 · $406,843

## Abstract

Project Summary
Red tide blooms significantly cause coastal ecosystem damage and a series of environmental diseases on
people who live nearby the coast or eat the red tide contaminated seafoods. Despite their ecological and
environmental significance, research in mechanistically understanding the red tide bloom occurrence is limited.
Plasma membrane G protein-coupled receptor (GPCRs) are widely implicated in multi-cellular eukaryotic signal
transductions, but their roles in the unicellular eukaryotic organism have not been well-explored. Recent studies
implied the involvements of primitive GPCRs in the bioluminescent pathway of dinoflagellates, including
Lingulodinium polyedra. L. polyedra is a prolific single-celled dinoflagellate implicated in red tide events and
highly sensitive to wave turbulence and predator behavior. Gaq-type heterotrimeric G proteins, one of major
signal pathways in the GPCR signaling, have been extensively reported for their associations with
mechanochemical transducer. Our preliminary bioinformatic analysis indicate a set of L. polyedra GPCRs are
implied in the bioluminescent pathway. The top 5 receptor candidates (RC1 through RC5) were subjected to
anti-sense DNA oligonucleotide knockdown experiments. The knockdowns indicate that RC3 is a promising
candidate involved in the bioluminescent signaling. Based on our preliminary data, this R21 project will further
explore the role of RC3 in dinoflagellate L. polyedra bioluminescence, starting from confirmation its shear force-
sensitivity, molecularly determining its role in bioluminescent signaling and biophysically characterizing the
receptor and related signaling partners. The project’s three specific aims include the following: Aim 1, RT-qPCR
will be used for evaluating the mRNA levels of RC3 and its effectors in the signaling pathway. CRISPR/Cas9
knockout strategy will also be used for validating the role of RC3 in shear-force elicited bioluminescence signaling.
In Aim 2, we will first heterogeneously express RC3 in a yeast-based GPCR preparation system and then use
monobromobiamine-based florescent spectroscopy to study its conformational responses to shear-forces and
copepodamide lipids. In Aim 3, we will determine whether RC3 related bioluminescence signal pathway is Gaq-
dependent, using both knockdown and knockout experiments along with in vitro measurements, including
GTPase-GloTM assay and [3H]GDP based GDP release assay. The R21 project data will determine if the
dinoflagellate derived GPCR—RC3 is associated with shear force-induced bioluminescence. The project’s
completion will establish a priming foundation to completely study the molecular mechanism of GPCR signaling
in the red tide blooms, with the far-reaching significance of controlling or modulating bloom occurrences.

## Key facts

- **NIH application ID:** 10708533
- **Project number:** 1R21ES035378-01
- **Recipient organization:** UNIVERSITY OF SOUTH FLORIDA
- **Principal Investigator:** Libin Ye
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $406,843
- **Award type:** 1
- **Project period:** 2023-08-23 → 2026-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10708533

## Citation

> US National Institutes of Health, RePORTER application 10708533, The Role of G Protein-coupled Receptors in Red Tide Dinoflagellate Bioluminescence (1R21ES035378-01). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10708533. Licensed CC0.

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