Structure, turnover, and heme-mediated suppression of the level of mRNA encoding rat liver delta-aminolevulinate synthase

J Biol Chem. 1988 Nov 5;263(31):15973-9.

Abstract

lambda gt11 cDNA libraries were constructed with poly(A)+ RNA preparations from both porphyric chicken and rat livers. A cDNA which encodes chicken hepatic delta-aminolevulinate synthase was cloned by screening with an anti-chicken liver delta-aminolevulinate synthase antibody. Using this cDNA as a probe, cDNAs encoding the entire protein coding sequence of rat hepatic delta-aminolevulinate synthase were then cloned. The complete nucleotide sequences of the cDNAs have been determined. The result predicts that the rat hepatic pre-delta-aminolevulinate synthase comprises 642 amino acids. We measured the half-life of the hepatic delta-aminolevulinate synthase mRNA by RNA blot hybridization analysis using allylisopropylacetamide-induced porphyric rats as an experimental model and the rat cDNA as a hybridization probe. The half-life of the mRNA determined by the injection of alpha-amanitin is as short as 20 min. This value is significantly shorter than the estimated half-lives of most other mRNAs in the differentiated tissues of animals. The effect of hemin administration on the level of hepatic delta-amino-levulinate synthase mRNA was also examined. The half-disappearance time of the mRNA after the hemin administration was essentially the same as that determined by alpha-amanitin or actinomycin D, and no additive effect was observed between alpha-amanitin and hemin on the half-life determination. The results provide convincing evidence that heme inhibits the transcription of delta-aminolevulinate synthase mRNA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 5-Aminolevulinate Synthetase / genetics*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Genes / drug effects*
  • Heme / pharmacology*
  • Liver / enzymology*
  • Molecular Sequence Data
  • RNA, Messenger / drug effects
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Rats
  • Suppression, Genetic / drug effects*

Substances

  • RNA, Messenger
  • Heme
  • 5-Aminolevulinate Synthetase

Associated data

  • GENBANK/J04044