Identification and characterization of up-regulated genes during chondrocyte hypertrophy.
Chondrocyte hypertrophy involves de novo acquisition and/or increased expression of certain gene products including, among others, type X collagen, alkaline phosphatase, and matrix metalloproteinases. To analyze further the genetic program associated with chondrocyte hypertrophy, we have employed a modification of the polymerase chain reaction-mediated subtractive hybridization method of Wang and Brown (Wang and Brown [1991] Proc. Natl. Acad. Sci 88:11505). Cultures of hypertrophic tibial chondrocytes and nonhypertrophic sternal cells were used for poly A+ RNA isolation. Among 50 individual cDNA fragments isolated for up-regulated hypertrophic genes, 18 were tentatively identified by their similarities to entries in the GenBank database, whereas the other 32 showed no significant similarity. The identified genes included translational and transcriptional regulatory factors, ribosomal proteins, the enzymes transglutaminase and glycogen phosphorylase, type X collagen (highly specific for hypertrophic cartilage matrix), gelsolin, and the carbohydrate-binding protein galectin. Two of these, transglutaminase and galectin, were cloned and were further characterized. The chondrocyte transglutaminase revealed previously in hypertrophic cartilage by immunochemical methods appears to be the chicken equivalent of mammalian factor XIIIa (showing 75% overall protein similarity). The chicken chondrocyte galectin is a variant of mammalian galectin-3. Galectins are known to bind to components found in hypertrophic cartilage, and factor XIIIa is known to crosslink some of the same components, possibly modifying them for calcification and/or removal.
Department of Anatomy and Cellular Biology, Tufts University Health Science Schools, Boston, Massachusetts 02111, USA.
M
M
Nurminskaya
T F
TF
Linsenmayer
D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002642:Chick Embryo; D018076:DNA, Complementary / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis* / Q000235:genetics; D005326:Fetal Proteins / Q000096:biosynthesis* / Q000235:genetics; D037502:Galectin 3; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D006132:Growth Plate / Q000166:cytology / Q000254:growth & development*; D008969:Molecular Sequence Data; D009924:Organ Culture Techniques; D010012:Osteogenesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013249:Sternum / Q000166:cytology; D013382:Subtraction Technique; D013977:Tibia; D011503:Transglutaminases / Q000096:biosynthesis / Q000235:genetics
D000943:Antigens, Differentiation; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D005326:Fetal Proteins; D037502:Galectin 3; D012333:RNA, Messenger; D011503:Transglutaminases