Essential alterations of heparan sulfate during the differentiation of embryonic stem cells to Sox1-enhanced green fluorescent protein-expressing neural progenitor cells.
Embryonic stem (ES) cells can be cultured in conditions that either maintain pluripotency or allow differentiation to the three embryonic germ layers. Heparan sulfate (HS), a highly polymorphic glycosaminoglycan, is a critical cell surface coreceptor in embryogenesis, and in this paper we describe its structural transition from an unusually low-sulfated variant in ES cells to a more highly sulfated form in fluorescence-activated cell sorting-purified neural progenitor cells. The characteristic domain structure of HS was retained during this transformation. However, qualitative variations in surface sulfation patterns between ES and differentiated cells were revealed using HS epitope-specific antibodies and the HS-binding growth factor fibroblast growth factor 2 (FGF-2). Expression profiles of the HS modification enzymes indicated that both "early" (N-sulfotransferases) and "late" (6O- and 3O-sulfotransferases) sulfotransferases contributed to the alterations in sulfation patterning. An HS-null ES line was used to demonstrate the necessity for HS in neural differentiation. HS is a coreceptor for many of the protein effectors implicated in pluripotency and differentiation (e.g., members of the FGF family, bone morphogenic proteins, and fibronectin). We suggest that the stage-specific activities of these proteins are finely regulated by dynamic changes in sulfation motifs in HS chains. Disclosure of potential conflicts of interest is found at the end of this article.
Johnson
Claire E
CE
Department of Medical Oncology, Cancer Research UK, The University of Manchester, Manchester, UK.
Crawford
Brett E
BE
Stavridis
Marios
M
Ten Dam
Gerdy
G
Wat
Annie L
AL
Rushton
Graham
G
Ward
Christopher M
CM
Wilson
Valerie
V
van Kuppevelt
Toin H
TH
Esko
Jeffrey D
JD
Smith
Austin
A
Gallagher
John T
JT
Merry
Catherine L R
CL
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006497:Heparan Sulfate / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors; D013431:Sulfates / Q000378:metabolism; D014162:Transfection; D014404:Tubulin / Q000378:metabolism
D004268:DNA-Binding Proteins; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; D006497:Heparan Sulfate; D006609:High Mobility Group Proteins; D011993:Recombinant Fusion Proteins; D055748:SOXB1 Transcription Factors; D013431:Sulfates; D014404:Tubulin; C098693:Sox1 protein, mouse; C462941:beta3 tubulin, mouse; C485184:enhanced green fluorescent protein