Structurally-discovered KLF4 variants accelerate and stabilize reprogramming to pluripotency

Autor: Takaaki Sato, Yuri An, Mami Matsuo-Takasaki, Dorian Luijkx, Tohru Terada, Koji Hisatake, Jingyue Li, Eiji Sugihara, Dan Song, Shiho Aizawa, Ken Nishimura, Yohei Hayashi, Akihiro Kuno, Miho Takami, Evgeniia Borisova, Fumiaki Yumoto
Rok vydání: 2022
Předmět:
Zdroj: iScience, Vol 25, Iss 1, Pp 103525-(2022)
iScience
ISSN: 2589-0042
Popis: Summary Non-genetically modified somatic cells can only be inefficiently and stochastically reprogrammed to pluripotency by exogenous expression of reprogramming factors. Low competence of natural reprogramming factors may prevent the majority of cells to successfully and synchronously reprogram. Here we screened DNA-interacting amino acid residues in the zinc-finger domain of KLF4 for enhanced reprogramming efficiency using alanine-substitution scanning methods. Identified KLF4 L507A mutant accelerated and stabilized reprogramming to pluripotency in both mouse and human somatic cells. By testing all the variants of L507 position, variants with smaller amino acid residues in the KLF4 L507 position showed higher reprogramming efficiency. L507A bound more to promoters or enhancers of pluripotency genes, such as KLF5, and drove gene expression of these genes during reprogramming. Molecular dynamics simulations predicted that L507A formed additional interactions with DNA. Our study demonstrates how modifications in amino acid residues of DNA-binding domains enable next-generation reprogramming technology with engineered reprogramming factors.
Graphical abstract
Highlights • KLF4 L507A variant accelerates and stabilizes reprogramming to pluripotency • KLF4 L507A has distinctive features of transcriptional binding and activation • KLF4 L507A may acquire a unique conformation with additional DNA interaction • Smaller amino acid residues in L507 position cause higher reprogramming efficiency
Stem cell plasticity; Molecular biology
Databáze: OpenAIRE
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