title: Elf5-centered transcription factor hub controls trophoblast stem cell self-renewal and differentiation through stoichiometry-sensitive shifts in target gene networks.
abstract: Elf5 is a transcription factor with pivotal roles in the trophoblast compartment, where it reinforces a trophoblast stem cell (TSC)-specific transcriptional circuit. However, Elf5 is also present in differentiating trophoblast cells that have ceased to express other TSC genes such as Cdx2 and Eomes. In the present study, we aimed to elucidate the context-dependent role of Elf5 at the interface between TSC self-renewal and the onset of differentiation. We demonstrate that precise levels of Elf5 are critical for normal expansion of the TSC compartment and embryonic survival, as Elf5 overexpression triggers precocious trophoblast differentiation. Through integration of protein interactome, transcriptome, and genome-wide chromatin immunoprecipitation data, we reveal that this abundance-dependent function is mediated through a shift in preferred Elf5-binding partners; in TSCs, Elf5 interaction with Eomes recruits Tfap2c to triply occupied sites at TSC-specific genes, driving their expression. In contrast, the Elf5 and Tfap2c interaction becomes predominant as their protein levels increase. This triggers binding to double- and single-occupancy sites that harbor the cognate Tfap2c motif, causing activation of the associated differentiation-promoting genes. These data place Elf5 at the center of a stoichiometry-sensitive transcriptional network, where it acts as a molecular switch governing the balance between TSC proliferation and differentiation.
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, United Kingdom;
authors.forename: Paulina A
authors.initials: PA
authors.lastname: Latos
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, United Kingdom;
authors.forename: Arnold R
authors.initials: AR
authors.lastname: Sienerth
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, United Kingdom;
authors.forename: Alexander
authors.initials: A
authors.lastname: Murray
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, United Kingdom;
authors.forename: Claire E
authors.initials: CE
authors.lastname: Senner
authors.affiliation: Graduate School of Pharmaceutical Sciences, Animal Resource Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan;
authors.forename: Masanaga
authors.initials: M
authors.lastname: Muto
authors.affiliation: Graduate School of Pharmaceutical Sciences, Animal Resource Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan;
authors.forename: Masahito
authors.initials: M
authors.lastname: Ikawa
authors.affiliation: Proteomics Group, The Babraham Institute, Cambridge CB22 3AT, United Kingdom;
authors.forename: David
authors.initials: D
authors.lastname: Oxley
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom;
authors.forename: Sarah
authors.initials: S
authors.lastname: Burge
authors.affiliation: Department of Physiology, Faculty of Medicine, University of Toronto, Toronto, Ontario M5S 1A8, Canada; Department of Obstetrics and Gynaecology, Faculty of Medicine, University of Toronto, Toronto, Ontario M5G 1E2, Canada.
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Cox
authors.affiliation: Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, United Kingdom;
authors.forename: Myriam
authors.initials: M
authors.lastname: Hemberger
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D000066673:Cell Self Renewal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D053263:Gene Regulatory Networks; D051379:Mice; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C500759:Elf5 protein, mouse; D014157:Transcription Factors
keywords: control of differentiation; self-renewal; transcription factor complexes; transcriptional networks; trophoblast stem cells