Characterization and hepatic expression of rat alpha 1-inhibitor III mRNA

A 2.5-kilobase cDNA clone (AF7), encoding 785 amino acids, was isolated from a rat liver cDNA library constructed in the expression vector lambda gt11. M13 vector sequence analysis yielded a deduced protein primary structure that was 89% homologous to the prototype alpha 1-inhibitor III (alpha 1I3) sequence presented in the preceding paper by Braciak et al. (Braciak, T. A., Northemann, W., Hudson, G. O., Shields, B. R., Gehring, M. R., and Fey, G. H. (1988) J. Biol. Chem. 263, 3999-4012) with regard to exact matches and 92% homologous when considering chemically conserved residues. The clone also possessed 100% homology to the putative bait region of a variant clone (pRLA1I3/27J) of alpha 1I3. Such sequence data demonstrates that the AF7 clone corresponds to a member of the family of variant alpha 1I3 mRNAs. Furthermore, this report presents the entire mRNA sequence corresponding to the 3'-half of alpha 1I3 variant 27J. We have utilized AF7 cDNA to study the expression of alpha 1I3 messenger RNA encoding this liver-specific glycoprotein under conditions known to alter hepatic gene expression. Our data reveal that alpha 1I3 mRNA expression is not only regulated during the acute-phase response but is also modulated in response to a variety of changing physiological conditions, most notably liver development. Steady state levels of mRNA were quantified using Northern blot techniques and laser densitometry. During acute phase response initiated by turpentine injection, the relative abundance of alpha 1I3 mRNA decreased 4-5-fold over a period of 24 h. Following partial hepatectomy, the regenerating liver expressed six-fold less alpha 1I3 mRNA than untreated liver after 24 h. This reduced level was maintained over a 2-day period. We have also demonstrated that alpha 1I3 mRNA expression is developmentally regulated. Fetal rat liver did not contain detectable concentrations of rat alpha 1I3 mRNA even as late as 4 days prior to birth. However, trace amounts were observed from birth until approximately 20 days of age when alpha 1I3 mRNA levels increased 10-fold to maximal adult quantities over the following 2 or 3 weeks. During the course of pregnancy, alpha 1I3 mRNA remained essentially constant until approximately 4 days prior to birth when a precipitous decline to 40% of the original level was noted. Subsequently, normal values were gradually restored over a 30-day postpartum period.(ABSTRACT TRUNCATED AT 400 WORDS)