Characterization of a cDNA for chicken osteopontin: expression during bone development, osteoblast differentiation, and tissue distribution.
The chicken bone phosphoprotein (approximately 66-kDa BPP) is a major noncollagenous component of bone and is the major phosphoprotein synthesized by cultured chicken embryo osteoblasts [Gotoh, Y., Gerstenfeld, L. C., & Glimcher, M. J. (1990) Eur. J. Biochem. 87, 49-58]. A cDNA clone for this protein was isolated from an expression library made from embryonic chicken bone mRNA. The complete primary protein sequence of 264 amino acids was deduced from the cDNA sequence inclusive of a 16 amino acid signal peptide sequence and terminated by 4 in-frame stop sequences. A sequence alignment indicated an approximate 35% overall similarity in protein sequence between the avian approximately 66-kDa BPP and the mammalian protein osteopontin, while at the nucleotide level 60% similarity was observed. Features of this sequence which showed the greatest similarity to mammalian osteopontin included a region in which seven of nine consecutive residues are aspartic acid, a recognition sequence for integrin-mediated cell binding (-Arg-Gly-Asp), and four possible recognition sequences for phosphorylation by casein kinase II. Hybridization analysis indicated a message of 1.5 kb found predominantly in bone and kidney. The mRNA was inducible in phorbol ester treated primary cultures of chondrocytes which show no expression under normal growth conditions. A temporal induction was seen during osteoblastic differentiation both in vivo and in vitro, thus suggesting that regulation of the approximately 66-kDa BPP is under transcriptional control during osteoblast development. In summary, both the protein's primary structure and its biological features suggest that it is the avian homologue to mammalian protein osteopontin.
Moore
M A
MA
Department of Orthopaedic Research, Harvard Medical School, Boston, Massachusetts.
Gotoh
Y
Y
Rafidi
K
K
Gerstenfeld
L C
LC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001846:Bone Development; D002454:Cell Differentiation; D002642:Chick Embryo; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D053495:Osteopontin; D010750:Phosphoproteins / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D012795:Sialoglycoproteins / Q000235:genetics*
D010750:Phosphoproteins; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D053495:Osteopontin; D004247:DNA