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An interaction between Gβγ and RNA polymerase II regulates transcription in cardiac fibroblasts

ABSTRACT: An interaction between Gβγ and RNA polymerase II regulates transcription in cardiac fibroblasts

PROVIDER: PRJNA614512 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11394450.fastq.gz	Fastqsanger.gz
	SRR11394451.fastq.gz	Fastqsanger.gz
	SRR11394452.fastq.gz	Fastqsanger.gz
	SRR11394453.fastq.gz	Fastqsanger.gz
	SRR11394454.fastq.gz	Fastqsanger.gz

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An interaction between Gβγ and RNA polymerase II regulates transcription in cardiac fibroblasts

Project description:Gβγ subunits are involved in an array of different signalling processes in various compartments of the cell, including the nucleus. To gain further insight into the functions of nuclear Gβγ, we investigated the functional role of Gβγ signalling in the regulation of GPCR-mediated gene expression in primary rat neonatal cardiac fibroblasts. Here, we demonstrate that following activation of the angiotensin II type I receptor in these cells, Gβγ dimers interact with RNA polymerase II (RNAPII). Our findings suggest that Gβ1 recruitment to RNAPII negatively regulates the fibrotic transcriptional response, which can be overcome by strong fibrotic stimuli. The Gβγ-RNAPII interaction was also investigated in HEK 293 cells and was regulated by signaling pathways that diverged from those operating in cardiac fibroblasts, suggesting that although it may be a conserved feature of transcriptional regulation, such regulation may be cell-specific. Taken together, our studies reveal a novel interaction between Gβγ subunits and RNA polymerase II and expand the role for Gβγ signalling in cardiac fibrosis.

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Project description:Gβγ subunits are involved in many different signalling processes in various compartments of the cell, including the nucleus. To gain insight into the functions of nuclear Gβγ, we investigated the functional role of Gβγ signalling in regulation of GPCR-mediated gene expression in primary rat neonatal cardiac fibroblasts. Following activation of the angiotensin II type I receptor in these cells, Gβγ dimers interact with RNA polymerase II (RNAPII). Our findings suggest that Gβ1γ recruitment to RNAPII negatively regulates the fibrotic transcriptional response, which can be overcome by strong fibrotic stimuli. In these specific proteomics experiments, we compared the differential protein abundance in Gβ1 knockdown and WT cardiac fibroblasts following angiotensin II treatment using a bottom-up data-dependent approach.

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Project description:Hematopoietic mutations in epigenetic regulators like DNA methyltransferase 3 alpha (DNMT3A) drive clonal hematopoiesis of indeterminate potential (CHIP) and are associated with adverse prognosis in patients with heart failure (HF). The interactions between CHIP-mutated cells and other cardiac cell types remain unknown. Here, we identify fibroblasts as potential interaction partners of CHIP-mutated monocytes using combined transcriptomic data from peripheral blood mononuclear cells of HF patients with and without CHIP and cardiac tissue. We demonstrate that DNMT3A inactivation augments macrophage-to-cardiac fibroblasts interactions and induces cardiac fibrosis in mice and humans. Mechanistically, DNMT3A inactivation increases the release of heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) to activate cardiac fibroblasts. These findings not only identify a novel pathway of DNMT3A CHIP-driver mutation-induced instigation and progression of HF, but may also provide a rationale for the development of new anti-fibrotic strategies.

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