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ATP is dispensable for both miRNA- and Smaug-mediated deadenylation reactions

Sho Niinuma1,2, Yukihide Tomari1,2

  • 1Institute of Molecular and Cellular Biosciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan.

RNA (New York, N.Y.)
|March 3, 2017
PubMed

Insights

Adenosine triphosphate (ATP) is not required for microRNA (miRNA)- or Smaug-induced deadenylation. Instead, accumulated adenosine monophosphate (AMP) inhibits this process, highlighting the need for caution with ATP depletion methods.

Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biochemistry

Background:

  • MicroRNAs (miRNAs) and Smaug protein recruit the CCR4-NOT deadenylase complex to shorten poly(A) tails.
  • ATP was previously thought to be essential for miRNA- or Smaug-mediated deadenylation due to observed inhibition upon ATP depletion.

Purpose of the Study:

  • To investigate the role of ATP in miRNA- and Smaug-induced deadenylation.
  • To clarify the mechanism behind the observed inhibition of deadenylation in the presence of ATP depletion.

Main Methods:

  • Experiments were conducted using Drosophila cell lysate.
  • The study monitored deadenylation activity in the presence and absence of an ATP-regenerating system.
  • Adenosine monophosphate (AMP) levels and deadenylase activity were assessed.

Main Results:

  • In the absence of an ATP-regenerating system, ATP was rapidly consumed to AMP.
  • AMP was identified as a potent inhibitor of deadenylation.
  • Hydrolysis of AMP restored deadenylation activity, indicating AMP accumulation was the inhibitory factor.

Conclusions:

  • ATP is dispensable for deadenylation induced by miRNAs or Smaug.
  • AMP accumulation, not ATP depletion, inhibits the deadenylation reaction.
  • Caution is advised when using ATP depletion methods to study deadenylation.
Keywords:
ATPCCR4-NOT complexSmaugdeadenylationmicroRNA

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