Expression of translation initiation factor IF2 is regulated during osteoblast differentiation.
We isolated and characterized a cDNA for the N-terminal half of the eukaryotic initiation of translation factor 2 (cIF2) during a screen of chicken osteoblast cDNAs. The apparent size of the message for this protein, approximately 5.6 kb, is slightly larger in size than that for human IF2 (hIF2). There is a high degree of sequence similarity between the human and chicken N-terminal portions of the protein that extends to the encoding nucleotide sequence. The tissue specific expression pattern for cIF2 and hIF2 are similar, being moderately abundant in brain, liver, and skeletal muscle, and detectable in kidney, chondrocytes, and freshly isolated osteoblasts. The ratio of message for cIF2 to that of beta-actin was 0.10 and 0.18 for liver and brain. Message levels peak in osteoblasts between 8 and 12 days of culture, coinciding with high levels of matrix protein synthesis. At peak expression, the ratio of cIF2:beta-actin for 8 day osteoblasts was 0.76. Treatment of osteoblast cultures with cycloheximide markedly reduces the level of cIF2 message indicating that novel protein synthesis is required for its expression. Hybridization of RNA samples from either chicken osteoblasts or a human osteoblast cell line with a probe for a subunit of human eukaryotic initiation of translation factor 2 (eIF2alpha), the housekeeping initiation factor, indicates that levels of eIF2 remain low. With hIF2, cIF2 represents the only other vertebrate homolog of IF2 for which a major portion of the coding sequence has been identified. This is the first report of regulated expression for a eukaryotic IF2 and is the first demonstration of its abundance in osteoblasts.
Weber
J A
JA
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802,USA.
Gay
C V
CV
D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002645:Chickens; D017124:Conserved Sequence / Q000235:genetics; D018076:DNA, Complementary / Q000302:isolation & purification; D015852:Eukaryotic Initiation Factor-2 / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism*; D010452:Peptide Biosynthesis / Q000502:physiology; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism
D018076:DNA, Complementary; D015852:Eukaryotic Initiation Factor-2; D012313:RNA