Over expression of wild type or a catalytically dead mutant of Sirtuin 6 does not influence NFκB responses.
SIRT6 is involved in inflammation, aging and metabolism potentially by modulating the functions of both NFκB and HIF1α. Since it is possible to make small molecule activators and inhibitors of Sirtuins we wished to establish biochemical and cellular assays both to assist in drug discovery efforts and to validate whether SIRT6 represents a valid drug target for these indications. We confirmed in cellular assays that SIRT6 can deacetylate acetylated-histone H3 lysine 9 (H3K9Ac), however this deacetylase activity is unusually low in biochemical assays. In an effort to develop alternative assay formats we observed that SIRT6 overexpression had no influence on TNFα induced nuclear translocation of NFκB, nor did it have an effect on nuclear mobility of RelA/p65. In an effort to identify a gene expression profile that could be used to identify a SIRT6 readout we conducted genome-wide expression studies. We observed that overexpression of SIRT6 had little influence on NFκB-dependent genes, but overexpression of the catalytically inactive mutant affected gene expression in developmental pathways.
Platform Technology Sciences, GlaxoSmithKline, Gunnels Wood Road, Stevenage, Hertfordshire, United Kingdom.
Rachel
R
Grimley
Oxana
O
Polyakova
Jessica
J
Vamathevan
Joanne
J
McKenary
Brian
B
Hayes
Champa
C
Patel
Janet
J
Smith
Angela
A
Bridges
Andrew
A
Fosberry
Anshu
A
Bhardwaja
Bernadette
B
Mouzon
Chun-Wa
CW
Chung
Nathalie
N
Barrett
Nicola
N
Richmond
Sundip
S
Modha
Roberto
R
Solari
D000107:Acetylation; D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006657:Histones / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism*; D037761:Sirtuins / Q000235:genetics* / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D006657:Histones; D016328:NF-kappa B; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; C516592:SIRT6 protein, human; D037761:Sirtuins