Unknown

Dataset Information

0

Regulation of heme oxygenase-1 protein expression by miR-377 in combination with miR-217.


ABSTRACT: Heme oxygenase-1 (HO-1) enzyme plays a critical role in metabolizing the excess heme generated during hemolysis. Our previous studies suggested that during intravascular hemolysis the expression of HO-1 protein is not sufficient to reduce the oxidative burden of free heme in the vasculature. This led us to hypothesize that a post-translational mechanism of control exists for HO-1 expression. Micro-RNAs (miRNA) affect gene expression by post-transcriptional gene regulation of transcripts. We performed in silico analysis for the human HMOX1-3' untranslated region (3' UTR) and identified candidate miRNA binding sites. Two candidate miRNAs, miR-377 and miR-217, were cloned and co-transfected with a luciferase vector containing the human HMOX1-3'UTR region. The combination of miR-377 and miR-217 produced a 58% reduction in HMOX1-3'UTR luciferase reporter expression compared with controls. The same constructs were then used to assess how overexpression of miR-217 and miR-377 affected HO-1 levels after induction with hemin. Cells transfected with the combination of miR-377 and miR-217 exhibited no change in HMOX1 mRNA levels, but a significant reduction in HMOX1 (HO-1) protein expression and enzyme activity compared with non-transfected hemin-stimulated controls. Transfection with either miR-377 or miR-217 alone did not produce a significant decrease in HO-1 protein expression or enzyme activity. Knockdown of miR-217 and miR-377 in combination leads to up-regulation of HO-1 protein. Exposure to hemin induced a significant reduction in miR-217 expression and a trend toward decreased miR-377 expression in two different cells lines. In summary, these data suggests that the combination of miR-377 and miR-217 help regulate HO-1 protein expression in the presence of hemin.

SUBMITTER: Beckman JD 

PROVIDER: S-EPMC3030323 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature

altmetric image

Publications

Regulation of heme oxygenase-1 protein expression by miR-377 in combination with miR-217.

Beckman Joan D JD   Chen Chunseng C   Nguyen Julia J   Thayanithy Venugopal V   Subramanian Subbaya S   Steer Clifford J CJ   Vercellotti Gregory M GM  

The Journal of biological chemistry 20101124 5


Heme oxygenase-1 (HO-1) enzyme plays a critical role in metabolizing the excess heme generated during hemolysis. Our previous studies suggested that during intravascular hemolysis the expression of HO-1 protein is not sufficient to reduce the oxidative burden of free heme in the vasculature. This led us to hypothesize that a post-translational mechanism of control exists for HO-1 expression. Micro-RNAs (miRNA) affect gene expression by post-transcriptional gene regulation of transcripts. We perf  ...[more]

Similar Datasets

| S-EPMC5437967 | biostudies-literature
| S-EPMC7377845 | biostudies-literature
| S-EPMC3788786 | biostudies-literature
| S-EPMC4561190 | biostudies-literature
| S-EPMC6760526 | biostudies-literature
| S-EPMC2781329 | biostudies-other
| S-EPMC5316511 | biostudies-literature
| S-EPMC5553546 | biostudies-literature
| S-EPMC3748626 | biostudies-literature
| S-EPMC8270315 | biostudies-literature