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Structure and expression of dog apolipoprotein C-II and C-III mRNAs. Implications for the evolution and functional
Abstract:
The complete cDNAs for canine apolipoprotein (apo) C-II and C-III have been cloned in the plasmid vector, pUC-18, and the phage vector, lambda gt11. With 32P-labeled nick-translated cDNAs as hybridization probes, apoC-II and apoC-III mRNAs were found to be expressed in canine liver, jejunum, ileum, colon, pancreas, spleen, ovary, and bladder at the following relative concentrations: 100, 35, 30, 10, 15, 5, 30, and 2% for apoC-II, and 100, 35, 15, 2, 20, 5, 30, and 5% for apoC-III. Neither mRNA was detected in canine brain, kidney, lung, muscle, oviduct, uterus, or testis. Both mRNAs are approximately 700 nucleotides in length. Complete sequence analysis revealed that apoC-II mRNA codes for a protein of 101 amino acid residues including 22 residues in the signal peptide and 79 residues in the mature peptide region. ApoC-III mRNA codes for a protein of 100 amino acid residues, including 20 in the signal peptide and 80 residues in the mature peptide region. It contains a single cysteine residue in position 54. Secondary structural analysis shows the presence of two regions each of amphipathic helix for apoC-II (residues 14-33 and 44-63) and apoC-III (residues 16-33 and 50-69). Computer analysis of the amphipathic helical regions demonstrates the importance of variation in the dihedral angles of the side chains in the charge microenvironment of the helices. Statistical analysis of sequence similarities between human apoC-II, apoC-III, apoA-II, apoA-I, apoE, and apoA-IV, and dog apoC-II and apoC-III, and rat apoC-III, apoA-II, apoA-I, apoE, and apoA-IV indicates that all these proteins have evolved rapidly, especially in the rat in which apoC-III has evolved at three times the rate in man and dog. Further, the shorter apolipoproteins, apoA-II, apoC-II, and a-poC-III, have evolved more rapidly than the longer ones, apoA-I, apoE, and apoA-IV. Some regions in the individual apolipoproteins are better conserved than others. The rate of evolution of individual regions seems to be related to the stringency of their functional requirements.