title: LncRNA-HIT Functions as an Epigenetic Regulator of Chondrogenesis through Its Recruitment of p100/CBP Complexes.
abstract: Gene expression profiling in E 11 mouse embryos identified high expression of the long noncoding RNA (lncRNA), LNCRNA-HIT in the undifferentiated limb mesenchyme, gut, and developing genital tubercle. In the limb mesenchyme, LncRNA-HIT was found to be retained in the nucleus, forming a complex with p100 and CBP. Analysis of the genome-wide distribution of LncRNA-HIT-p100/CBP complexes by ChIRP-seq revealed LncRNA-HIT associated peaks at multiple loci in the murine genome. Ontological analysis of the genes contacted by LncRNA-HIT-p100/CBP complexes indicate a primary role for these loci in chondrogenic differentiation. Functional analysis using siRNA-mediated reductions in LncRNA-HIT or p100 transcripts revealed a significant decrease in expression of many of the LncRNA-HIT-associated loci. LncRNA-HIT siRNA treatments also impacted the ability of the limb mesenchyme to form cartilage, reducing mesenchymal cell condensation and the formation of cartilage nodules. Mechanistically the LncRNA-HIT siRNA treatments impacted pro-chondrogenic gene expression by reducing H3K27ac or p100 activity, confirming that LncRNA-HIT is essential for chondrogenic differentiation in the limb mesenchyme. Taken together, these findings reveal a fundamental epigenetic mechanism functioning during early limb development, using LncRNA-HIT and its associated proteins to promote the expression of multiple genes whose products are necessary for the formation of cartilage.
authors.affiliation: Skeletal Biology Program, Shriners Hospitals for Children, Portland, Oregon, United States of America.
authors.forename: Hanqian L
authors.initials: HL
authors.lastname: Carlson
authors.affiliation: Program in Epithelial Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Quinn
authors.affiliation: Program in Epithelial Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Yul W
authors.initials: YW
authors.lastname: Yang
authors.affiliation: Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, United States of America.
authors.forename: Chelsea K
authors.initials: CK
authors.lastname: Thornburg
authors.affiliation: Program in Epithelial Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, California, United States of America.
authors.forename: Howard Y
authors.initials: HY
authors.lastname: Chang
authors.affiliation: Skeletal Biology Program, Shriners Hospitals for Children, Portland, Oregon, United States of America.|Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, Oregon, United States of America.
authors.forename: H Scott
authors.initials: HS
authors.lastname: Stadler
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D020219:Chondrogenesis / Q000235:genetics*; D044127:Epigenesis, Genetic / Q000235:genetics; D005121:Extremities / Q000254:growth & development; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D018878:Limb Buds / Q000254:growth & development; D008648:Mesoderm / Q000254:growth & development; D051379:Mice; D062085:RNA, Long Noncoding / Q000096:biosynthesis / Q000235:genetics*; D020729:p120 GTPase Activating Protein / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D062085:RNA, Long Noncoding; D020729:p120 GTPase Activating Protein