title: SUMOylation Regulates Growth Factor Independence 1 in Transcriptional Control and Hematopoiesis.
abstract: Cell fate specification requires precise coordination of transcription factors and their regulators to achieve fidelity and flexibility in lineage allocation. The transcriptional repressor growth factor independence 1 (GFI1) is comprised of conserved Snail/Slug/Gfi1 (SNAG) and zinc finger motifs separated by a linker region poorly conserved with GFI1B, its closest homolog. Moreover, GFI1 and GFI1B coordinate distinct developmental fates in hematopoiesis, suggesting that their functional differences may derive from structures within their linkers. We show a binding interface between the GFI1 linker and the SP-RING domain of PIAS3, an E3-SUMO (small ubiquitin-related modifier) ligase. The PIAS3 binding region in GFI1 contains a conserved type I SUMOylation consensus element, centered on lysine-239 (K239). In silico prediction algorithms identify K239 as the only high-probability site for SUMO modification. We show that GFI1 is modified by SUMO at K239. SUMOylation-resistant derivatives of GFI1 fail to complement Gfi1 depletion phenotypes in zebrafish primitive erythropoiesis and granulocytic differentiation in cultured human cells. LSD1/CoREST recruitment and MYC repression by GFI1 are profoundly impaired for SUMOylation-resistant GFI1 derivatives, while enforced expression of MYC blocks granulocytic differentiation. These findings suggest that SUMOylation within the GFI1 linker favors LSD1/CoREST recruitment and MYC repression to govern hematopoietic differentiation.
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Andrade
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Velinder
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Singer
authors.affiliation: Department of Pediatrics, University of Utah School of Medicine, Salt Lake City, Utah, USA Primary Children's Hospital, Salt Lake City, Utah, USA.
authors.forename: Luke
authors.initials: L
authors.lastname: Maese
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Diana
authors.initials: D
authors.lastname: Bareyan
authors.affiliation: Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Nguyen
authors.affiliation: Department of Radiation Oncology, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Mahesh B
authors.initials: MB
authors.lastname: Chandrasekharan
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: Helena
authors.initials: H
authors.lastname: Lucente
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: David
authors.initials: D
authors.lastname: McClellan
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA.
authors.forename: David
authors.initials: D
authors.lastname: Jones
authors.affiliation: Department of Medicine, University of Utah School of Medicine, Salt Lake City, Utah, USA Center for Investigational Therapeutics, Huntsman Cancer Institute, Salt Lake City, Utah, USA.
authors.forename: Sunil
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Susan Lehman Cullman Laboratory for Cancer Research, Department of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers Cancer Institute of New Jersey, Rutgers, The State University of New Jersey, Piscataway, New Jersey, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Oncological Sciences, University of Utah School of Medicine, Salt Lake City, Utah, USA Department of Pediatrics, University of Utah School of Medicine, Salt Lake City, Utah, USA Primary Children's Hospital, Salt Lake City, Utah, USA Nuclear Control Program, Huntsman Cancer Institute, Salt Lake City, Utah, USA michael.engel@hci.utah.edu.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Engel
mesh_terms: D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002454:Cell Differentiation; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D018922:HL-60 Cells; D006410:Hematopoiesis; D056466:Histone Demethylases / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism; D051379:Mice; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D041681:NIH 3T3 Cells; D011485:Protein Binding; D050828:Protein Inhibitors of Activated STAT / Q000737:chemistry / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D058207:Sumoylation
chemical_list: C111977:GFI1B protein, human; C489427:MYC protein, human; D018832:Molecular Chaperones; C484745:PIAS3 protein, human; D050828:Protein Inhibitors of Activated STAT; D011518:Proto-Oncogene Proteins; D016271:Proto-Oncogene Proteins c-myc; D012097:Repressor Proteins; D056466:Histone Demethylases; C495195:KDM1A protein, human; D008239:Lysine