RRC ID 67197
Author Jaiswal PK, Aljebali L, Gaumond MH, Oh CD, Yasuda H, Moffatt P.
Title Biochemical characteristics of the chondrocyte-enriched SNORC protein and its transcriptional regulation by SOX9.
Journal Sci Rep
Abstract Snorc (Small NOvel Rich in Cartilage) has been identified as a chondrocyte-specific gene in the mouse. Yet little is known about the SNORC protein biochemical properties, and mechanistically how the gene is regulated transcriptionally in a tissue-specific manner. The goals of the present study were to shed light on those important aspects. The chondrocyte nature of Snorc expression was confirmed in mouse and rat tissues, in differentiated (day 7) ATDC5, and in RCS cells where it was constitutive. Topological mapping and biochemical analysis brought experimental evidences that SNORC is a type I protein carrying a chondroitin sulfate (CS) attached to serine 44. The anomalous migration of SNORC on SDS-PAGE was due to its primary polypeptide features, suggesting no additional post-translational modifications apart from the CS glycosaminoglycan. A highly conserved SOX9-binding enhancer located in intron 1 was necessary to drive transcription of Snorc in the mouse, rat, and human. The enhancer was active independently of orientation and whether located in a heterologous promoter or intron. Crispr-mediated inactivation of the enhancer in RCS cells caused reduction of Snorc. Transgenic mice carrying the intronic multimerized enhancer drove high expression of a βGeo reporter in chondrocytes, but not in the hypertrophic zone. Altogether these data confirmed the chondrocyte-specific nature of Snorc and revealed dependency on the intronic enhancer binding of SOX9 for transcription.
Volume 10(1)
Pages 7790
Published 2020-5-8
DOI 10.1038/s41598-020-64640-x
PII 10.1038/s41598-020-64640-x
PMID 32385306
PMC PMC7210984
MeSH Amino Acid Sequence Animals Base Sequence Cell Line Cells, Cultured Chondrocytes / metabolism* Chondroitin Sulfates / metabolism Gene Expression Regulation* Humans Membrane Proteins / chemistry Membrane Proteins / genetics* Membrane Proteins / metabolism* Mice Mice, Transgenic Open Reading Frames Protein Binding Protein Domains Proteoglycans / chemistry Proteoglycans / genetics* Proteoglycans / metabolism* Rats SOX9 Transcription Factor / metabolism* Transcription Initiation Site Transcription, Genetic
IF 3.998
Human and Animal Cells ATDC5(RCB0565)