title: Natural antisense transcript for hyaluronan synthase 2 (HAS2-AS1) induces transcription of HAS2 via protein O-GlcNAcylation.
abstract: Changes in the microenvironment organization within vascular walls are critical events in the pathogenesis of vascular pathologies, including atherosclerosis and restenosis. Hyaluronan (HA) accumulation into artery walls supports vessel thickening and is involved in many cardiocirculatory diseases. Excessive cytosolic glucose can enter the hexosamine biosynthetic pathway, increase UDP-N-acetylglucosamine (UDP-GlcNAc) availability, and lead to modification of cytosolic proteins via O-linked attachment of the monosaccharide β-N-GlcNAc (O-GlcNAcylation) from UDP-GlcNAc by the enzyme O-GlcNAc transferase. As many cytoplasmic and nuclear proteins can be glycosylated by O-GlcNAc, we studied whether the expression of the HA synthases that synthesize HA could be controlled by O-GlcNAcylation in human aortic smooth muscle cells. Among the three HAS isoenzymes, only HAS2 mRNA increased after O-GlcNAcylation induced by glucosamine treatments or by inhibiting O-GlcNAc transferase with PUGNAC (O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate). We found that the natural antisense transcript of HAS2 (HAS2-AS1) was absolutely necessary to induce the transcription of the HAS2 gene. Moreover, we found that O-GlcNAcylation modulated HAS2-AS1 promoter activation by recruiting the NF-κB subunit p65, but not the HAS2 promoter, whereas HAS2-AS1 natural antisense transcript, working in cis, regulated HAS2 transcription by altering the chromatin structure around the HAS2 proximal promoter via O-GlcNAcylation and acetylation. These results indicate that HAS2 transcription can be finely regulated not only by recruiting transcription factors to the promoter as previously described but also by modulating chromatin accessibility by epigenetic modifications.
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Vigetti
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Deleonibus
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Moretto
authors.affiliation: Department of Nephrology, Cardiff University School of Medicine, Heath Park, Cardiff CF14 4XN, United Kingdom.
authors.forename: Timothy
authors.initials: T
authors.lastname: Bowen
authors.affiliation: Institute of Pharmacology and Clinical Pharmacology, University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, 40225 Düsseldorf, Germany.
authors.forename: Jens W
authors.initials: JW
authors.lastname: Fischer
authors.affiliation: Institute of Pharmacology and Clinical Pharmacology, University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, 40225 Düsseldorf, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Grandoch
authors.affiliation: Department of Vascular and Endovascular Surgery, University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Oberhuber
authors.affiliation: Laboratory of Cell Biochemistry and Biology, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Dona C
authors.initials: DC
authors.lastname: Love
authors.affiliation: Laboratory of Cell Biochemistry and Biology, NIDDK, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: John A
authors.initials: JA
authors.lastname: Hanover
authors.affiliation: Dipertimento di Biotecnologia e Scienze della Vita, Università degli Studi dell'Insubria, 21100 Varese, Italy, and.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Cinquetti
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Karousou
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Viola
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: D'Angelo
authors.affiliation: Biomedical Engineering ND20, The Cleveland Clinic, Cleveland, Ohio 44195.
authors.forename: Vincent C
authors.initials: VC
authors.lastname: Hascall
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: De Luca
authors.affiliation: From the Dipartimento di Scienze Chirurgiche e Morfologiche, Università degli Studi dell'Insubria, Varese, Italy, alberto.passi@uninsubria.it.
authors.forename: Alberto
authors.initials: A
authors.lastname: Passi
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry; D000818:Animals; D001011:Aorta / Q000201:enzymology; D001483:Base Sequence; D002467:Cell Nucleus / Q000201:enzymology; D002843:Chromatin / Q000737:chemistry; D003593:Cytoplasm / Q000201:enzymology; D044127:Epigenesis, Genetic; D015971:Gene Expression Regulation, Enzymologic; D020868:Gene Silencing; D014453:Glucuronosyltransferase / Q000235:genetics* / Q000502:physiology; D006801:Humans; D000076002:Hyaluronan Synthases; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry; D032389:Myocytes, Smooth Muscle / Q000201:enzymology; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D009005:Monosaccharides; D012333:RNA, Messenger; D017351:N-Acetylglucosaminyltransferases; C449880:O-GlcNAc transferase; D014453:Glucuronosyltransferase; C101754:HAS2 protein, human; C491099:Has2 protein, mouse; D000076002:Hyaluronan Synthases; D000117:Acetylglucosamine
keywords: Epigenetics; Glycobiology; Glycosaminoglycan; Glycosylation; Hyaluronan; Long Noncoding RNA (Long ncRNA, lncRNA); Proteoglycan