Heparan sulfate 6-o-sulfotransferase is essential for muscle development in zebrafish.
Heparan sulfate proteoglycans function in development and disease. They consist of a core protein with attached heparan sulfate chains that are altered by a series of carbohydrate-modifying enzymes and sulfotransferases. Here, we report on the identification and characterization of a gene encoding zebrafish heparan sulfate 6-O-sulfotransferase (hs6st) that shows high homology to other heparan sulfate 6-O-sulfotransferases. When expressed as a fusion protein in cultured cells, the protein shows specific 6-O-sulfotransferase activity and preferentially acts on the iduronosyl N-sulfoglycosamine. In the developing embryo, hs6st is expressed in the brain, the somites, and the fins; the same structures that were affected upon morpholino-mediated functional knockdown. Morpholino injections significantly inhibited 6-O- but not 2-O-sulfation as assessed by HPLC. Morphants display disturbed somite specification independent of the somite oscillator mechanism and have impaired muscle differentiation. In conclusion, our results show that transfer of sulfate to specific positions on glycosaminoglycans is essential for muscle development.
Bink
Robert J
RJ
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands.
Habuchi
Hiroko
H
Lele
Zsolt
Z
Dolk
Edward
E
Joore
Jos
J
Rauch
Gerd-Jörg
GJ
Geisler
Robert
R
Wilson
Stephen W
SW
den Hertog
Jeroen
J
Kimata
Koji
K
Zivkovic
Danica
D
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000166:cytology / Q000196:embryology* / Q000201:enzymology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010641:Phenotype; D019170:Somites / Q000201:enzymology; D015238:Sulfotransferases / Q000235:genetics* / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics*
D019812:Heparan Sulfate Proteoglycans; D016376:Oligonucleotides, Antisense; D015238:Sulfotransferases; C404189:heparan sulfate 6-O-sulfotransferase