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Elife. 2019 Jan 24;8. pii: e42341. doi: 10.7554/eLife.42341.

Exploration of CTCF post-translation modifications uncovers Serine-224 phosphorylation by PLK1 at pericentric regions during the G2/M transition.

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Department of Molecular Biology, Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, United States.
Department of Genetics, Harvard Medical School, Boston, United States.
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States.
Department of Molecular Biology, Massachusetts General Hospital, Boston, United States.
Cell Signaling Technology, Danvers, United States.
Contributed equally


The zinc finger CCCTC-binding protein (CTCF) carries out many functions in the cell. Although previous studies sought to explain CTCF multivalency based on sequence composition of binding sites, few examined how CTCF post-translational modification (PTM) could contribute to function. Here, we performed CTCF mass spectrometry, identified a novel phosphorylation site at Serine 224 (Ser224-P), and demonstrate that phosphorylation is carried out by Polo-like kinase 1 (PLK1). CTCF Ser224-P is chromatin-associated, mapping to at least a subset of known CTCF sites. CTCF Ser224-P accumulates during the G2/M transition of the cell cycle and is enriched at pericentric regions. The phospho-obviation mutant, S224A, appeared normal. However, the phospho-mimic mutant, S224E, is detrimental to mouse embryonic stem cell colonies. While ploidy and chromatin architecture appear unaffected, S224E mutants differentially express hundreds of genes, including p53 and p21. We have thus identified a new CTCF PTM and provided evidence of biological function.


CTCF; PLK1; cell cycle; chromosome architecture; chromosomes; gene expression; mouse; post-translational modification

Conflict of interest statement

BD, AK, AD, AA, FW, RS No competing interests declared, CF Employee of Cell Signaling Technologies. There are no other competing interests to declare. JL Reviewing editor, eLife, and is a cofounder and member of the Scientific Advisory Boards of Translate Bio and Fulcrum Therapeutics.

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