title: Molecular analysis of heparan sulfate biosynthetic enzyme machinery and characterization of heparan sulfate structure in Nematostella vectensis.
abstract: HS (heparan sulfate) proteoglycans are key regulators of vital processes in the body. HS chains with distinct sequences bind to various protein ligands, such as growth factors and morphogens, and thereby function as important regulators of protein gradient formation and signal transduction. HS is synthesized through the concerted action of many different ER (endoplasmic reticulum) and Golgi-resident enzymes. In higher organisms, many of these enzymes occur in multiple isoforms that differ in substrate specificity and spatial and temporal expression. In order to investigate how the structural complexity of HS has evolved, in the present study we focused on the starlet sea anemone (Nematostella vectensis), which belongs to the Anthozoa, which are considered to have retained many ancestral features. Members of all of the enzyme families involved in the generation and modification of HS were identified in Nematostella. Our results show that the enzymes are highly conserved throughout evolution, but the number of isoforms varies. Furthermore, the HS polymerases [Ext (exostosin) enzymes Ext1, Ext2 and Ext-like3] represent distinct subgroups, indicating that these three genes have already been present in the last common ancestor of Cnidaria and Bilateria. In situ hybridization showed up-regulation of certain enzymes in specific areas of the embryo at different developmental stages. The specific mRNA expression pattern of particular HS enzymes implies that they may play a specific role in HS modifications during larval development. Finally, biochemical analysis of Nematostella HS demonstrates that the sea anemone synthesizes a polysaccharide with a unique structure.
authors.affiliation: Department of Biomedicine, University of Bergen, Jonas Lies vei 91, Bergen, Norway.
authors.forename: Almir
authors.initials: A
authors.lastname: Feta
authors.forename: Anh-Tri
authors.initials: AT
authors.lastname: Do
authors.forename: Fabian
authors.initials: F
authors.lastname: Rentzsch
authors.forename: Ulrich
authors.initials: U
authors.lastname: Technau
authors.forename: Marion
authors.initials: M
authors.lastname: Kusche-Gullberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000737:chemistry; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006497:Heparan Sulfate / Q000096:biosynthesis* / Q000737:chemistry*; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012615:Sea Anemones / Q000196:embryology / Q000201:enzymology*; D016415:Sequence Alignment
chemical_list: D004187:Disaccharides; D006497:Heparan Sulfate; D017351:N-Acetylglucosaminyltransferases; D012333:RNA, Messenger