@article{a77da93ec91040fca544ab8a941546df,
title = "EKLF/Klf1 regulates erythroid transcription by its pioneering activity and selective control of RNA Pol II pause-release",
abstract = "EKLF/Klf1 is a zinc-finger transcription activator essential for erythroid lineage commitment and terminal differentiation. Using ChIP-seq, we investigate EKLF DNA binding and transcription activation mechanisms during mouse embryonic erythropoiesis. We utilize the Nan/+ mouse that expresses the EKLF-E339D (Nan) variant mutated in its conserved zinc-finger region and address the mechanism of hypomorphic and neomorphic changes in downstream gene expression. First, we show that Nan-EKLF limits normal EKLF binding to a subset of its sites. Second, we find that ectopic binding of Nan-EKLF occurs largely at enhancers and activates transcription through pioneering activity. Third, we find that for a subset of ectopic targets, gene activation is achieved in Nan/+ only by Nan-EKLF binding to distal enhancers, leading to RNA polymerase II pause-release. These results have general applicability to understanding how a DNA binding variant factor confers dominant disruptive effects on downstream gene expression even in the presence of its normal counterpart.",
keywords = "CBP, CP: Molecular biology, ChIP-seq, EKLF/Klf1, H3K27ac, Nan, RNA polymerase, elongation, erythropoiesis, pausing, pioneering, transcription",
author = "Kaustav Mukherjee and Bieker, {James J.}",
note = "Funding Information: This work was supported by a Black Family Stem Cell Institute postdoctoral award to K.M. and by NIH grant R01 DK046865 to J.J.B. We thank Mohan Dangeti for initial RNA Pol II experiments, Chrysoula Deligianni for ATAC-seq, Zelda Salfati at the MSSM Genomics Core for advice on NGS library optimization, Tom Moran and J. Andrew Duty of the MSSM Hybridoma Core Facility for establishing the 7B2a/IgG2a clones, and the MSSM Flow Cytometry Core facility. We thank Cheryl A. Keller and Ross Hardison for advice on the ATAC-seq protocol. We acknowledge Nithya Gnanapragasam, Sanjana Pillay, and Antanas Planutis for ongoing discussions; Sree Chinta for RNA analysis; and Li Xue for mouse husbandry. Funding Information: This work was supported by a Black Family Stem Cell Institute postdoctoral award to K.M. and by NIH grant R01 DK046865 to J.J.B. We thank Mohan Dangeti for initial RNA Pol II experiments, Chrysoula Deligianni for ATAC-seq, Zelda Salfati at the MSSM Genomics Core for advice on NGS library optimization, Tom Moran and J. Andrew Duty of the MSSM Hybridoma Core Facility for establishing the 7B2a/IgG2a clones, and the MSSM Flow Cytometry Core facility. We thank Cheryl A. Keller and Ross Hardison for advice on the ATAC-seq protocol. We acknowledge Nithya Gnanapragasam, Sanjana Pillay, and Antanas Planutis for ongoing discussions; Sree Chinta for RNA analysis; and Li Xue for mouse husbandry. K.M. performed experiments and analyzed the data; K.M. and J.J.B. designed the experiments, interpreted the results, and wrote the paper. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = dec,
day = "20",
doi = "10.1016/j.celrep.2022.111830",
language = "English",
volume = "41",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "12",
}