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20250306_Antarctic_Ulva_Metabolomics

ABSTRACT: The algal holobiont, which consists of algae and the bacteria living on it, has developed strategies to handle environmental stresses, such as changes in temperature. The green alga Ulva (Chlorophyta) is home to bacteria that help the algae grow and develop, from germlings to mature plants. These bacteria release chemicals like thallusin, N-acyl homoserine lactone, and ectoine, which support the health and growth of the algae. However, there is limited research on the metabolome of cold-adapted Ulva holobionts, especially those from hard-to-reach regions like Antarctica, and how they respond to environmental changes. In this study, we examine the metabolites found in Ulva and its associated bacteria from Antarctica (Potter Cove) using high-resolution mass spectrometry. We identified small polar and zwitterionic metabolites, such as cysteinolic acid, ectoine, glutamine, glycerol, and proline, in the surface tissues of the algae with MALDI-MS. By profiling these metabolites using UHPLC-HRMS and ZIC-HILIC chromatography, we found that their levels change significantly in response to heat-stressed Antarctic Ulva holobionts. Our results suggest that environmental change can affect the interaction between seaweeds and their microbes by altering the metabolites involved in their chemical exchanges. The Data contains the Ulva metabolome profiles (.mzXML) analyzed using ZIC-HILIC chromatography coupled with HRMS (QExactive). Acquisition was conducted with the polarity switch mode and ddMS acquisition was done on the QC pool samples. The Metadata, normalization factor and results files from CD 3.3 can be found in the supplementary files.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Ulva (ncbitaxon:3118)

SUBMITTER: Dr. Marine Vallet

PROVIDER: MSV000097253 | MassIVE | Thu Mar 06 15:39:00 GMT 2025

REPOSITORIES: MassIVE

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Project description:The family of Zic transcription factors (TFs) are required for cerebellar development, and their binding is highly enriched at developmentally regulated enhancers active in cerebellar granule neurons at the progenitor stage around postnatal day 7 (P7) and the mature adult stage at P60 in the mouse cerebellum. Interestingly, Zic1/2 ChIP-seq data reveal that the Zics change their binding profile over time. To characterize these differences in Zic targeting between early and late cerebellar maturation, we compared the sequence underlying Zic Peaks, their relationship with active chromatin and gene expression at P7 and P60. From this we found Zic peaks tend to be large and bind open-active chromatin. Strikingly, Zic peaks shift from binding at promoter proximal regions at P7 to distal enhancer regions at P60. We hypothesize that other TFs collaborate with the Zic TFs to change their binding targets and affect their regulatory activity. A multi-tiered approach was used to predict TF binding at those Zic ChIP sites. We assessed motif enrichment (HOMER) and in-vivo TF binding profiles (BART) to determine putative TFs collaborating with Zic to drive this regulation. We then validated the presence of the predicted TFs in granule neurons using gene expression. This workflow identified known and novel distinct collaborators of Zic between early and late development. Early collaborates of Zic includes bHLH factors and chromatin remodelers whereas late collaborators of Zic are activity regulated TFs which are markers of synaptic maturation. To identify Zic’s developmental gene targets in cerebellar maturation, we used H3K4me3 PLAC-seq data, which captures promoter-enhancer loops from the adult mouse cerebellum and Hi-C data from the young mouse cerebellum to map genes to Zic bound enhancers. This revealed Zic as a transcriptional activator of late developmental genes in the cerebellum. Using this same approach with in vitro Zic KD, we were able to identify Zic dependent developmental gene targets which have functions in axonogenesis, axon guidance, and ion channel signaling, which are integral in proper maturation of a neuron. These integrated analyses reveal how Zic and other TFs regulate temporal expression of CGN developmental genes and provides information that will enhance our understanding of the molecular mechanisms that regulate the mode of TF function at enhancers.

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20250306_Antarctic_Ulva_Metabolomics

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