Human heme oxygenase cDNA and induction of its mRNA by hemin

Authors


Correspondence to S. Shibahara, Friedrich-Miescher-Institut, P.O. Box 2543, CH-4002 Basel, Switzerland

Abstract

Hemin treatment increased both activity and mRNA level of heme oxygenase in human macrophages. Using poly(A)-rich RNA prepared from human macrophages treated with hemin, we have constructed a cDNA library in the Okayama-Berg vector. The human heme oxygenase cDNA was isolated by screening this library with a rat cDNA and was subjected to nucleotide sequence analysis. The deduced human heme oxygenase is composed of 288 amino acids with a molecular mass of 32800 Da. The homology in amino acid sequences between rat and human heme oxygenase is 80%. Like rat heme oxygenase, human enzyme has a putative membrane segment at its carboxyl terminus, which is probably essential for the insertion of heme oxygenase into endoplasmic reticulum. Both rat and human heme oxygenase have no cysteine residues. Recently we have shown that rat heme oxygenase is a heat-shock protein [J. Biol. Chem. 262, 12889–12892 (1987)], and therefore we examined the effects of heat treatment on the induction of heme oxygenase in human macrophages and glioma cells. In contrast to hemin treatment, heat treatment had no apparent effects in either human cell line on the activity of heme oxygenase and its mRNA levels. These results suggest that human heme oxygenase may not be a heat-shock protein.

Abbreviation
DMEM

Dulbecco's minimal essential medium

Enzymes
 

Alkaline phosphatase or orthophosphoric-monoester phosphohydrolase (alkaline optimum) (EC 3.1.3.1)

 

DNA ligase or poly(deoxyribonucleotide):poly(deoxyribonucleotide) ligase (AMP-forming, NMN-forming) (EC 6.5.1.2)

 

DNA polymerase I or deoxynucleoside-triphosphate: DNA deoxynucleotidyltransferase (EC 2.7.7.7)

 

endoribonuclease H (EC 3.1.26.4)

 

heme oxygenase (EC 1.14.99.3)

 

polynucleotide 5′-hydroxyl-kinase or ATP: 5′-dephosphonucleotide 5′-phosphotransferase (EC 2.7.1.78)

 

reverse transcriptase or deoxynucleoside triphosphate: DNA deoxynucleotidyltransferase (RNA-directed)(EC 2.7.7.49)

 

terminal deoxyribonucleotidyl transferase or nucleoside-triphosphate:DNA deoxynucleotidylexotransferase (EC 2.7.7.31)

Ancillary