title: Conversion of Sox17 into a pluripotency reprogramming factor by reengineering its association with Oct4 on DNA.
abstract: Very few proteins are capable to induce pluripotent stem (iPS) cells and their biochemical uniqueness remains unexplained. For example, Sox2 cooperates with other transcription factors to generate iPS cells, but Sox17, despite binding to similar DNA sequences, cannot. Here, we show that Sox2 and Sox17 exhibit inverse heterodimerization preferences with Oct4 on the canonical versus a newly identified compressed sox/oct motif. We can swap the cooperativity profiles of Sox2 and Sox17 by exchanging single amino acids at the Oct4 interaction interface resulting in Sox2KE and Sox17EK proteins. The reengineered Sox17EK now promotes reprogramming of somatic cells to iPS, whereas Sox2KE has lost this potential. Consistently, when Sox2KE is overexpressed in embryonic stem cells it forces endoderm differentiation similar to wild-type Sox17. Together, we demonstrate that strategic point mutations that facilitate Sox/Oct4 dimer formation on variant DNA motifs lead to a dramatic swap of the bioactivities of Sox2 and Sox17.
authors.affiliation: Laboratory for Structural Biochemistry and Genome Institute of Singapore, Singapore, Singapore.
authors.forename: Ralf
authors.initials: R
authors.lastname: Jauch
authors.forename: Irene
authors.initials: I
authors.lastname: Aksoy
authors.forename: Andrew Paul
authors.initials: AP
authors.lastname: Hutchins
authors.forename: Calista Keow Leng
authors.initials: CK
authors.lastname: Ng
authors.forename: Xian Feng
authors.initials: XF
authors.lastname: Tian
authors.forename: Jiaxuan
authors.initials: J
authors.lastname: Chen
authors.forename: Paaventhan
authors.initials: P
authors.lastname: Palasingam
authors.forename: Paul
authors.initials: P
authors.lastname: Robson
authors.forename: Lawrence W
authors.initials: LW
authors.lastname: Stanton
authors.forename: Prasanna R
authors.initials: PR
authors.lastname: Kolatkar
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D003198:Computer Simulation; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D024002:HMGB Proteins / Q000235:genetics* / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D011485:Protein Binding / Q000235:genetics; D015202:Protein Engineering; D055503:Protein Multimerization; D055748:SOXB1 Transcription Factors / Q000235:genetics* / Q000378:metabolism; D055760:SOXF Transcription Factors / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D055748:SOXB1 Transcription Factors; D055760:SOXF Transcription Factors; C511606:Sox17 protein, mouse; C527783:Sox2 protein, mouse; D004247:DNA