Directed evolution of SIRT6 for improved deacylation and glucose homeostasis maintenance

Scientific Reports
Or GertmanAmir Aharoni

Abstract

Mammalian SIRT6 is a well-studied histone deacetylase that was recently shown to exhibit high protein deacylation activity enabling the removal of long chain fatty acyl groups from proteins. SIRT6 was shown to play key roles in cellular homeostasis by regulating a variety of cellular processes including DNA repair and glucose metabolism. However, the link between SIRT6 enzymatic activities and its cellular functions is not clear. Here, we utilized a directed enzyme evolution approach to generate SIRT6 mutants with improved deacylation activity. We found that while two mutants show increased deacylation activity at high substrate concentration and improved glucose metabolism they exhibit no improvement and even abolished deacetylation activity on H3K9Ac and H3K56Ac in cells. Our results demonstrate the separation of function between SIRT6 catalytic activities and suggest that SIRT6 deacylation activity in cells is important for glucose metabolism and can be mediated by still unknown acylated cellular proteins.

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Citations

Jan 30, 2020·ELife·Lior OnnDebra Toiber
Jun 11, 2020·The Journal of Biological Chemistry·Mark A Klein, John M Denu
Sep 22, 2020·Molecular Biology and Evolution·Adi HendlerAmir Aharoni
May 14, 2020·Nutrition Reviews·Ana Paula S CaldasHelen Hermana M Hermsdorff

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Methods Mentioned

BETA
deacylation
acetylation
ELISA
immunoprecipitation
PCR
transfection
flow cytometry
Assay

Software Mentioned

image J

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