Structure–function analysis of kdm2a

Access through your institution Buy or subscribe The authors generated a series of complexes of KDM2A bound to H3(A29-Y41)K36me2 peptides and grew crystals that diffracted to 1.75 Å

resolution. The complexes included various KDM2A mutants as well as peptide mutants; all were subsequently assayed _in vitro_ for demethylation activity. The authors found that KDM2A

recognizes three sequence context features that can explain its specificity to methylated H3K36: Gly33 and Gly34 residues at the −2 and −3 positions of the bound peptide (with Lys36 in

position 0), which occupy a narrow channel in KDM2A that cannot accommodate residues with side chains; a residue at the +2 position (Pro38) that can support peptide bending upon binding; and

an aromatic residue at the +5 position (Tyr41) that facilitates the insertion of the K36me2 side chain into the catalytic pocket of the enzyme. Mutating these residues, or mutating residues

in the enzyme that interact with them, severely hampered KDM2A activity. Consistent with this, other methylated Lys residues in H3 that are not targeted by KDM2A do not share these

recognition features. Methylated H3K36 has been implicated in cancer, so the authors went on to use HT1080 fibrosarcoma cells to examine the effects of expressing two structure-guided,

enzymatically inactive KDM2A mutants carrying mutations in the catalytic pocket. Overexpression of wild-type KDM2A but not of the mutants resulted in decreased H3K36me2 levels, whereas

H3K4me2 and H3K9me2 levels were unaffected. Conversely, depletion of KDM2A by RNAi led to an increase in H3K36me2, and complementation of these cells with RNAi-resistant wild-type KDM2A, but

not with RNAi-resistant mutant proteins, restored H3K36me2 levels. Importantly, KDM2A depletion caused chromosomal instability, evident by a marked increase in chromosome bridges and

micronuclei, and promoted anchorage-independent growth, which is a hallmark of cellular transformation. These phenotypes could also be rescued by wild-type but not by the catalytically

inactive mutants. Together, these results suggest that KDM2A may have tumour-suppressive functions. This is a preview of subscription content, access via your institution ACCESS OPTIONS

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institutional subscriptions * Read our FAQs * Contact customer support REFERENCES * Cheng, Z. et al. A molecular threading mechanism underlies Jumonji lysine demethylase KDM2A regulation of

methylated H3K36. _Genes Dev._ 28, 1758–1771 (2014) Article  CAS  PubMed  PubMed Central  Google Scholar  Download references Authors * Eytan Zlotorynski View author publications You can

also search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Zlotorynski, E. Structure–function analysis of KDM2A.

_Nat Rev Mol Cell Biol_ 15, 631 (2014). https://doi.org/10.1038/nrm3870 Download citation * Published: 28 August 2014 * Issue Date: October 2014 * DOI: https://doi.org/10.1038/nrm3870 SHARE

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