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Whole genome bisulfite sequencing of Picocholorum soloecismus across several days of its growth cycle

ABSTRACT: Epigenetic modifications have not been well characterized in most microalgae species, particularly those used for biofuel production. We report the first analysis of DNA methylation of the species Picocholorum soloecismus across its growth cycle using whole-genome bisulfite sequencing. DNA methylation occurs in all three cytosine contexts (CpG, CHH, CHG) in P. soloecismus. While global DNA methylation is low overall (~1.15% of the 15 MB genome), methylation occurs appreciable quantities (12.1%) in CpG dinucleotides in a bimodal distribution. The P. soloecismus genome becomes hypomethylated during the growth cycle (across several days in culture). 5-aza-2’-deoxycytidine (5AZA) treatment was used to alter DNA methylation patterns, which resulted in altered algal physiology and site specific changes in CpG DNA methylation.

ORGANISM(S): Picochlorum sp. 'soloecismus'

PROVIDER: GSE155500 | GEO | 2020/08/01

REPOSITORIES: GEO

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Project description:Atherosclerosis preferentially occurs in arterial regions of disturbed blood flow (d-flow), which alters gene expression, endothelial function, and atherosclerosis. Here, we show that d-flow regulates genome-wide DNA methylation patterns in a DNA methyltransferase (DNMT)-dependent manner. D-flow induced expression of DNMT1 in mouse arterial endothelium in vivo and in cultured endothelial cells by oscillatory shear (OS) in vitro. The DNMT inhibitor 5-Aza-2’deoxycytidine (5Aza) or DNMT1 siRNA significantly reduced OS-induced endothelial inflammation. Moreover, 5Aza reduced lesion formation in two ApoE-/- mouse atherosclerosis models. To identify the 5Aza mechanisms, we conducted two genome-wide studies: reduced representation bisulfite sequencing (RRBS) and microarray using endothelial-enriched gDNA and RNA, respectively, from the partially-ligated left carotid artery (LCA exposed to d-flow) and the right contralateral control (RCA) of mice treated with 5Aza or vehicle. Systems biological analyses using RRBS and transcriptome data revealed 11 mechanosensitive genes whose promoters were hypermethylated under d-flow conditions, but rescued by 5Aza treatment. Of those, the two transcription factors HoxA5 and Klf3 contain cAMP- response-elements, and their methylation status could serve as a mechanosensitive master switch in gene expression. Our results demonstrate that d-flow controls epigenomic DNA methylation patterns in a DNMT-dependent manner, which in turn alters endothelial gene expression and induces atherosclerosis. We used 6- to 8-week-old male C57Bl/6 mice (The Jackson Laboratory) according to the approved Institutional Animal Care and Use Committee protocol by Emory University. Mice were subjected to partial carotid ligation surgery under anesthesia. Brieﬂy, 3 of 4 caudal branches of LCA (left external carotid, internal carotid, and occipital artery) were ligated with 6-0 silk suture, although the superior thyroid artery was left intact. Development of low and oscillatory blood flow in the Left Carotid Artery of each mouse was determined by ultrasound measurements. Each sample contained total RNA from 3 pooled RCAs or LCAs. We ran 3 samples of LCA, RCA, AzaLCA, and AzaRCA on 2 microarrays.

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Whole genome bisulfite sequencing of Picocholorum soloecismus across several days of its growth cycle

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