title: The chromatin-binding protein HMGN3 stimulates histone acetylation and transcription across the Glyt1 gene.
abstract: HMGNs are nucleosome-binding proteins that alter the pattern of histone modifications and modulate the binding of linker histones to chromatin. The HMGN3 family member exists as two splice forms, HMGN3a which is full-length and HMGN3b which lacks the C-terminal RD (regulatory domain). In the present study, we have used the Glyt1 (glycine transporter 1) gene as a model system to investigate where HMGN proteins are bound across the locus in vivo, and to study how the two HMGN3 splice variants affect histone modifications and gene expression. We demonstrate that HMGN1, HMGN2, HMGN3a and HMGN3b are bound across the Glyt1 gene locus and surrounding regions, and are not enriched more highly at the promoter or putative enhancer. We conclude that the peaks of H3K4me3 (trimethylated Lys(4) of histone H3) and H3K9ac (acetylated Lys(9) of histone H3) at the active Glyt1a promoter do not play a major role in recruiting HMGN proteins. HMGN3a/b binding leads to increased H3K14 (Lys(14) of histone H3) acetylation and stimulates Glyt1a expression, but does not alter the levels of H3K4me3 or H3K9ac enrichment. Acetylation assays show that HMGN3a stimulates the ability of PCAF [p300/CREB (cAMP-response-element-binding protein)-binding protein-associated factor] to acetylate nucleosomal H3 in vitro, whereas HMGN3b does not. We propose a model where HMGN3a/b-stimulated H3K14 acetylation across the bodies of large genes such as Glyt1 can lead to more efficient transcription elongation and increased mRNA production.
authors.affiliation: Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK.
authors.forename: Gráinne
authors.initials: G
authors.lastname: Barkess
authors.forename: Yuri
authors.initials: Y
authors.lastname: Postnikov
authors.forename: Chrisanne D
authors.initials: CD
authors.lastname: Campos
authors.forename: Shivam
authors.initials: S
authors.lastname: Mishra
authors.forename: Gokula
authors.initials: G
authors.lastname: Mohan
authors.forename: Sakshi
authors.initials: S
authors.lastname: Verma
authors.forename: Michael
authors.initials: M
authors.lastname: Bustin
authors.forename: Katherine L
authors.initials: KL
authors.lastname: West
mesh_terms: D000107:Acetylation; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D050487:Glycine Plasma Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D024001:HMGN Proteins / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D014162:Transfection
chemical_list: D002843:Chromatin; D050487:Glycine Plasma Membrane Transport Proteins; D024001:HMGN Proteins; C532059:HMGN3 protein, mouse; D006657:Histones; C493267:Slc6a9 protein, mouse