| Structural highlights
Function
[ALKB5_HUMAN] Dioxygenase that demethylates RNA by oxidative demethylation: specifically demethylates N(6)-methyladenosine (m(6)A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes. Requires molecular oxygen, alpha-ketoglutarate and iron. Demethylation of m(6)A mRNA affects mRNA processing and export and is required for spermatogenesis.[1] [2]
Publication Abstract from PubMed
N6-Methyladenosine (m6A) is the most prevalent internal RNA modification in eukaryotes. ALKBH5 belongs to the AlkB family of dioxygenases and has been shown to specifically demethylate m6A in single stranded RNA. Here we report crystal structures of ALKBH5 in the presence of either its cofactors or the ALKBH5 inhibitor citrate. Catalytic assays demonstrate that the ALKBH5 catalytic domain can demethylate both ssRNA and ssDNA. We identify the tricarboxylic acid (TCA) cycle intermediate citrate as a modest inhibitor of ALKHB5 (IC50: ~488 muM). The structural analysis reveals that a loop region of ALKBH5 is immobilized by a disulfide bond which apparently excludes the binding of dsDNA to ALKBH5. We identify the m6A binding pocket of ALKBH5 and the key residues involved in m6A recognition using mutagenesis and ITC binding experiments.
Structures of human ALKBH5 demethylase reveal a unique binding mode for specific single stranded m6A RNA demethylation.,Xu C, Liu K, Tempel W, Demetriades M, Aik W, Schofield CJ, Min J J Biol Chem. 2014 Apr 28. PMID:24778178[3]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Thalhammer A, Bencokova Z, Poole R, Loenarz C, Adam J, O'Flaherty L, Schodel J, Mole D, Giaslakiotis K, Schofield CJ, Hammond EM, Ratcliffe PJ, Pollard PJ. Human AlkB homologue 5 is a nuclear 2-oxoglutarate dependent oxygenase and a direct target of hypoxia-inducible factor 1alpha (HIF-1alpha). PLoS One. 2011 Jan 14;6(1):e16210. doi: 10.1371/journal.pone.0016210. PMID:21264265 doi:https://dx.doi.org/10.1371/journal.pone.0016210
- ↑ Zheng G, Dahl JA, Niu Y, Fedorcsak P, Huang CM, Li CJ, Vagbo CB, Shi Y, Wang WL, Song SH, Lu Z, Bosmans RP, Dai Q, Hao YJ, Yang X, Zhao WM, Tong WM, Wang XJ, Bogdan F, Furu K, Fu Y, Jia G, Zhao X, Liu J, Krokan HE, Klungland A, Yang YG, He C. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol Cell. 2013 Jan 10;49(1):18-29. doi: 10.1016/j.molcel.2012.10.015. Epub 2012, Nov 21. PMID:23177736 doi:https://dx.doi.org/10.1016/j.molcel.2012.10.015
- ↑ Xu C, Liu K, Tempel W, Demetriades M, Aik W, Schofield CJ, Min J. Structures of human ALKBH5 demethylase reveal a unique binding mode for specific single stranded m6A RNA demethylation. J Biol Chem. 2014 Apr 28. PMID:24778178 doi:https://dx.doi.org/10.1074/jbc.M114.550350
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