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  • Title: Phenobarbital-mediated modulation of gene expression in rat liver. Analysis of cDNA clones.
    Author: Lechner MC, Sinogas C, Osório-Almeida ML, Freire MT, Chaumet-Riffaud P, Frain M, Sala-Trepat JM.
    Journal: Eur J Biochem; 1987 Mar 02; 163(2):231-8. PubMed ID: 3816806.
    Abstract:
    Phenobarbital evokes a pleiotypic response in the liver characterized by cell hypertrophy and mono-oxygenase induction. These phenomena arise through complex modulation mechanisms changing the pattern of protein synthesis, distinct from those triggered by other well known inducers, like steroid hormones or polycyclic hydrocarbons. To investigate the mechanisms involved in regulating the expression of the phenobarbital-inducible tissue-specific genes, we constructed two libraries of recombinant bacterial plasmids pBR322 in Escherichia coli. Each library contains cDNA copies of polysomal poly(A)-rich RNA obtained from control and 16-h phenobarbital-induced rat liver. A thousand cloned sequences from each library were screened by double-cross colony hybridization using [32P]cDNA prepared from homologous and heterologous poly(A)-rich RNAs as the probes. The statistical analysis of the results revealed that phenobarbital treatment significantly changes the relative abundance of different polysomal mRNA classes in rat liver. Clones corresponding to mRNAs clearly induced following phenobarbital treatment have been further selected by plasmid DNA dot hybridization, and used as probes for measuring the changes in each mRNA concentration in the whole cell and in the polysomal RNAs from rat livers, at different times after phenobarbital treatment. The fact that changes in the concentration of each specific mRNA in the polysomes does not parallel the variation of its total concentration in the cell indicates that the induced modulation of protein synthesis in the liver is brought about by mechanisms involving both transcriptional and translational regulation, since besides the increases in whole cellular mRNA concentration a marked mobilization of mRNA into active polysomes could be demonstrated during the onset of the adaptive response to phenobarbital.
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