RT Journal Article
SR Electronic
T1 Fe(III) heme sets an activation threshold for processing distinct groups of pri-miRNAs in mammalian cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.18.955294
DO 10.1101/2020.02.18.955294
A1 Weitz, Sara H.
A1 Quick-Cleveland, Jen
A1 Jacob, Jose P.
A1 Barr, Ian
A1 Senturia, Rachel
A1 Koyano, Kikuye
A1 Xiao, Xinshu
A1 Weiss, Shimon
A1 Guo, Feng
YR 2020
UL http://biorxiv.org/content/early/2020/02/19/2020.02.18.955294.abstract
AB The essential biological cofactor heme is synthesized in cells in the Fe(II) form. Oxidized Fe(III) heme is specifically required for processing primary transcripts of microRNAs (pri-miRNAs) by the RNA-binding protein DGCR8, a core component of the Microprocessor complex. It is unknown how readily available Fe(III) heme is in the largely reducing environment in human cells and how changes in cellular Fe(III) heme availability alter microRNA (miRNA) expression. Here we address the first question by characterizing DGCR8 mutants with various degrees of deficiency in heme-binding. We observed a strikingly simple correlation between Fe(III) heme affinity in vitro and the Microprocessor activity in HeLa cells, with the heme affinity threshold for activation estimated to be between 0.6-5 pM under typical cell culture conditions. The threshold is strongly influenced by cellular heme synthesis and uptake. We suggest that the threshold reflects a labile Fe(III) heme pool in cells. Based on our understanding of DGCR8 mutants, we reanalyzed recently reported miRNA sequencing data and conclude that heme is generally required for processing canonical pri-miRNAs, that heme modulates the specificity of Microprocessor, and that cellular heme level and differential DGCR8 heme occupancy alter the expression of distinct groups of miRNAs in a hierarchical fashion. Overall, our study provides the first glimpse of a labile Fe(III) heme pool important for a fundamental physiological function and reveal principles governing how Fe(III) heme modulates miRNA maturation at a genomic scale. We also discuss potential states and biological significance of the labile Fe(III) heme pool.