O-Linked β-N-Acetylglucosamine (O-GlcNAc) Regulates Emerin Binding to Barrier to Autointegration Factor (BAF) in a Chromatin- and Lamin B-enriched “Niche”

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Emerin, a membrane component of nuclear "lamina" networks with lamins and barrier to autointegration factor (BAF), is highly O -GlcNAc-modified (" O -GlcNAcylated") in mammalian cells. Mass spectrometry analysis revealed eight sites of O -GlcNAcylation, including Ser-53, Ser-54, Ser-87, Ser-171, and Ser-173. Emerin O -GlcNAcylation was reduced ∼50% by S53A or S54A mutation in vitro and in vivo . O -GlcNAcylation was reduced ∼66% by the triple S52A/S53A/S54A mutant, and S173A reduced O -GlcNAcylation of the S52A/S53A/S54A mutant by ∼30%, in vivo . We separated two populations of emerin, A-type lamins and BAF; one population solubilized easily, and the other required sonication and included histones and B-type lamins. Emerin and BAF associated only in histone- and lamin-B-containing fractions. The S173D mutation specifically and selectively reduced GFP-emerin association with BAF by 58% and also increased GFP-emerin hyper-phosphorylation. We conclude that β- N -acetylglucosaminyltransferase, an essential enzyme, controls two regions in emerin. The first region, defined by residues Ser-53 and Ser-54, flanks the LEM domain. O -GlcNAc modification at Ser-173, in the second region, is proposed to promote emerin association with BAF in the chromatin/lamin B "niche." These results reveal direct control of a conserved LEM domain nuclear lamina component by β- N -acetylglucosaminyltransferase, a nutrient sensor that regulates cell stress responses, mitosis, and epigenetics. Background: Nuclear membrane protein emerin binding to nuclear intermediate filaments (lamins) and BAF contributes to forming a nuclear "lamina" structure. Results: Emerin is O -GlcNAc-modified at eight sites: two (Ser-53 and Ser-54) influence further O -GlcNAcylation, and one (Ser-173) regulates association with BAF in the chromatin/lamin B "niche." Conclusion: O -GlcNAc transferase, a nutrient-responsive enzyme, regulates emerin. Significance: Emerin hyper- O -GlcNAcylation may contribute to cardiomyopathy and other conditions.