title: Hematopoietic Signaling Mechanism Revealed from a Stem/Progenitor Cell Cistrome.
abstract: Thousands of cis-elements in genomes are predicted to have vital functions. Although conservation, activity in surrogate assays, polymorphisms, and disease mutations provide functional clues, deletion from endogenous loci constitutes the gold-standard test. A GATA-2-binding, Gata2 intronic cis-element (+9.5) required for hematopoietic stem cell genesis in mice is mutated in a human immunodeficiency syndrome. Because +9.5 is the only cis-element known to mediate stem cell genesis, we devised a strategy to identify functionally comparable enhancers ("+9.5-like") genome-wide. Gene editing revealed +9.5-like activity to mediate GATA-2 occupancy, chromatin opening, and transcriptional activation. A +9.5-like element resided in Samd14, which encodes a protein of unknown function. Samd14 increased hematopoietic progenitor levels/activity and promoted signaling by a pathway vital for hematopoietic stem/progenitor cell regulation (stem cell factor/c-Kit), and c-Kit rescued Samd14 loss-of-function phenotypes. Thus, the hematopoietic stem/progenitor cell cistrome revealed a mediator of a signaling pathway that has broad importance for stem/progenitor cell biology.
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Hewitt
authors.affiliation: Institute for Basic Science, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea; Department of Chemistry, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: Duk Hyoung
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Prithvia
authors.initials: P
authors.lastname: Devadas
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Rajalekshmi
authors.initials: R
authors.lastname: Prathibha
authors.affiliation: Department of Biostatistics and Medical Informatics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA.
authors.forename: Chandler
authors.initials: C
authors.lastname: Zuo
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Rajendran
authors.initials: R
authors.lastname: Sanalkumar
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Kirby D
authors.initials: KD
authors.lastname: Johnson
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA.
authors.forename: Yoon-A
authors.initials: YA
authors.lastname: Kang
authors.affiliation: Institute for Basic Science, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea; Department of Chemistry, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-742, South Korea.
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Department of Biostatistics and Medical Informatics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA.
authors.forename: Colin N
authors.initials: CN
authors.lastname: Dewey
authors.affiliation: Department of Biostatistics and Medical Informatics, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA. Electronic address: keles@stat.wisc.edu.
authors.forename: Sunduz
authors.initials: S
authors.lastname: Keles
authors.affiliation: Department of Cell and Regenerative Biology, Carbone Cancer Center, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA; UW-Madison Blood Research Program, Madison, WI 53706, USA. Electronic address: ehbresni@wisc.edu.
authors.forename: Emery H
authors.initials: EH
authors.lastname: Bresnick
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D050989:GATA2 Transcription Factor / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction; D014158:Transcription, Genetic / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics*; D003598:Cytoskeletal Proteins
chemical_list: D050989:GATA2 Transcription Factor; D011506:Proteins; D019009:Proto-Oncogene Proteins c-kit; D034741:RNA, Small Interfering; C494712:Gata2 protein, mouse; C000601826:Samd14 protein, mouse; D003598:Cytoskeletal Proteins