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Application of MassSQUIRM for Quantitative Measurements of Lysine Demethylase Activity
07:02

Application of MassSQUIRM for Quantitative Measurements of Lysine Demethylase Activity

Published on: March 11, 2012

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SMYD3-mediated lysine methylation in the PH domain is critical for activation of AKT1

Yuichiro Yoshioka1,2, Takehiro Suzuki3, Yo Matsuo4

  • 1Section of Hematology/Oncology, Department of Medicine, The University of Chicago, MC2115 Chicago, IL 60637, USA.

Oncotarget
|September 15, 2016
PubMed

Insights

The protein lysine methyltransferase SMYD3 methylates AKT1 at lysine 14, a novel modification essential for AKT1 activation and cancer cell growth. This SMYD3-mediated methylation of AKT1 is a potential therapeutic target for anti-cancer treatments.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • AKT1 is a key serine/threonine kinase implicated in tumorigenesis and frequently overexpressed in cancers.
  • Post-translational modifications like phosphorylation and ubiquitination are known for AKT1, but methylation has not been previously reported.

Purpose of the Study:

  • To investigate the potential methylation of AKT1.
  • To identify the enzyme responsible for AKT1 methylation and elucidate its functional consequences in cancer.

Main Methods:

  • In vitro and in vivo methylation assays using protein lysine methyltransferase SMYD3.
  • Site-directed mutagenesis to substitute lysine 14 of AKT1.
  • Western blotting to assess AKT1 phosphorylation at threonine 308.
  • Cellular assays to evaluate plasma membrane accumulation and cancer cell growth rates.
  • SMYD3 knockdown and inhibitor treatments in cancer cells.

Main Results:

  • SMYD3 was identified as the enzyme that methylates lysine 14 (K14) in the PH domain of AKT1.
  • Methylation of AKT1 at K14 significantly reduced phosphorylation at threonine 308 (T308).
  • SMYD3 inhibition or knockdown attenuated AKT1 T308 phosphorylation in cancer cells.
  • K14 substitution diminished AKT1 plasma membrane localization and reduced cancer cell growth rates.

Conclusions:

  • SMYD3-mediated methylation of AKT1 at K14 is a critical regulatory mechanism for AKT1 activation.
  • This methylation event is essential for AKT1's role in promoting cancer cell proliferation.
  • SMYD3-mediated AKT1 methylation represents a promising therapeutic target for developing novel anti-cancer strategies.
Keywords:
AKT1PH domainSMYD3human cancerlysine methylation

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