John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, United States; Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, United States
David W Sant
John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, United States; Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, United States
Tyler C Huff
John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, United States; Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, United States
Sushmita Mustafi
John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, United States; Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, United States
Ryan K Muir
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
Allegra T Aron
Department of Chemistry, University of California, Berkeley, Berkeley, United States
Christopher J Chang
Department of Chemistry, University of California, Berkeley, Berkeley, United States; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States; Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States
Adam R Renslo
Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
Paula V Monje
The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, United States; Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, United States
John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, United States; Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Miller School of Medicine, Miami, United States; Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, United States; Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, United States
It is widely accepted that cAMP regulates gene transcription principally by activating the protein kinase A (PKA)-targeted transcription factors. Here, we show that cAMP enhances the generation of 5-hydroxymethylcytosine (5hmC) in multiple cell types. 5hmC is converted from 5-methylcytosine (5mC) by Tet methylcytosine dioxygenases, for which Fe(II) is an essential cofactor. The promotion of 5hmC was mediated by a prompt increase of the intracellular labile Fe(II) pool (LIP). cAMP enhanced the acidification of endosomes for Fe(II) release to the LIP likely through RapGEF2. The effect of cAMP on Fe(II) and 5hmC was confirmed by adenylate cyclase activators, phosphodiesterase inhibitors, and most notably by stimulation of G protein-coupled receptors (GPCR). The transcriptomic changes caused by cAMP occurred in concert with 5hmC elevation in differentially transcribed genes. Collectively, these data show a previously unrecognized regulation of gene transcription by GPCR-cAMP signaling through augmentation of the intracellular labile Fe(II) pool and DNA hydroxymethylation.
