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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
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A pre-microRNA-149 (miR-149) genetic variation affects miR-149 maturation and its ability to regulate the Puma

Su-Ling Ding1, Jian-Xun Wang, Jian-Qin Jiao

  • 1From the Division of Cardiovascular Research, State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.

The Journal of Biological Chemistry
|July 23, 2013
PubMed

Insights

Genetic variation in miR-149 precursor (rs71428439) impacts mature microRNA production and myocardial infarction risk. The G allele reduces mature miR-149, impairing its protective effects against apoptosis and mitochondrial fission.

Area of Science:

  • Genetics
  • Molecular Biology
  • Cardiovascular Research

Background:

  • MicroRNAs (miRNAs) are key gene regulators, but the impact of natural genetic variations in their precursors is not well understood.
  • Genetic variations within miRNA precursors can alter miRNA processing and function.
  • Understanding these variations is crucial for elucidating disease mechanisms.

Purpose of the Study:

  • To investigate the functional consequences of a specific polymorphism (rs71428439) in the Homo sapiens miR-149 precursor stem-loop region.
  • To determine the association of this polymorphism with myocardial infarction.
  • To elucidate the molecular mechanisms by which this polymorphism affects miR-149 function and cellular processes.

Main Methods:

  • Genotyping analysis of the A→G polymorphism (rs71428439) in myocardial infarction cases and controls.
  • Assessment of mature miR-149 production from different allelic precursors.
  • Functional assays examining miR-149's role in mitochondrial fission and apoptosis, including targeting of the p53-up-regulated modulator of apoptosis (Puma).
  • Evaluation of the polymorphism's effect on cardiac function in a mouse model of myocardial infarction.

Main Results:

  • The genotype distribution of the miR-149 polymorphism differed significantly between myocardial infarction patients and controls.
  • The G-allele precursor resulted in lower mature miR-149 production compared to the A-allele.
  • miR-149 inhibits mitochondrial fission and apoptosis by targeting Puma; the A to G variation attenuated this inhibitory effect.
  • The polymorphism influenced cardiac function in a mouse model of myocardial infarction.

Conclusions:

  • The A→G polymorphism in the miR-149 precursor (rs71428439) affects mature miR-149 biogenesis and function.
  • This polymorphism is associated with myocardial infarction and influences cellular responses to apoptotic stimuli.
  • The findings suggest a role for miR-149 precursor genetic variation in the pathogenesis of myocardial infarction, warranting further investigation.
Keywords:
Cardiovascular DiseaseGenetic PolymorphismHuman GeneticsMicroRNAMitochondrial Apoptosis

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