Cloning and sequence analysis of the rat augmenter of liver regeneration (ALR) gene: expression of biologically active recombinant ALR and demonstration of tissue distribution.
A full-length cDNA clone encoding a purified augmenter of liver regeneration (ALR) factor prepared from the cytosol of weanling rat livers was isolated. The 1.2-kb cDNA included a 299-bp 5' untranslated region, a 375-bp coding region, and a 550-bp 3' untranslated region. It encoded a protein consisting of 125 amino acids. The molecular weight of ALR calculated from the cDNA was 15,081, which is consistent with the size estimated by SDS/PAGE under reducing conditions. The molecular weight of the purified native ALR estimated by SDS/PAGE under nonreducing conditions was approximately 30,000; thus ALR apparently has a homodimeric structure. The recombinant ALR produced by expression of the cDNA in COS cells was tested in vivo in the canine Eck fistula model and found to have potency equivalent to the purified native ALR. The 125-aa sequence deduced from the rat ALR cDNA shows 50% homology to the amino acid sequence of the gene for oxidative phosphorylation and vegetative growth in the yeast Saccharomyces cerevisiae.
Hagiya
M
M
Pharmaceuticals Research Center, Toyobo Co., Ltd, Shiga, Japan.
Francavilla
A
A
Polimeno
L
L
Ihara
I
I
Sakai
H
H
Seki
T
T
Shimonishi
M
M
Porter
K A
KA
Starzl
T E
TE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D015870:Gene Expression; D006133:Growth Substances / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008099:Liver / Q000378:metabolism*; D008115:Liver Regeneration / Q000235:genetics; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010085:Oxidative Phosphorylation; D016133:Polymerase Chain Reaction; D011506:Proteins; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D050862:Oxidoreductases Acting on Sulfur Group Donors
D017931:DNA Primers; D018076:DNA, Complementary; D006133:Growth Substances; D011506:Proteins; D012333:RNA, Messenger; C088825:GFER protein, rat; D050862:Oxidoreductases Acting on Sulfur Group Donors