The sirtuins are
a family of genes encoding NAD(+)-dependent protein deacylases conserved in
several organisms including yeast, worms, flies, and mammals. This family of
genes has been shown to play crucial roles in the molecular pathways that
regulate cell survival, metabolism, and the diseases of aging. Because of the
diverse roles sirtuins play, research in the area of sirtuins has expanded into
multiple disciplines at an accelerated pace, identifying new and unexpected
roles for this family of genes. A diverse array of model organisms and
scientific techniques are used to study the sirtuins, which are presented in a
comprehensive manner in this volume of Methods in Molecular Biology. After
an introductory chapter, the first four chapters (Part I) focus on “Methods to generate
sirtuin biology tools,” one each for yeast, C . elegans, Drosophila
, and mammalian model organisms. In addition to reviewing common tools,
some techniques are presented along with these tools as examples for their use.
The next five chapters (Part II) focus on “Methods to identify sirtuin
substrates,” covering several enrichment and proteomic strategies to measure
changes in acylation of sirtuin substrates. The following five chapters (Part
III) cover “Methods to measure sirtuin activity,” which is one of the major
challenges currently facing the sirtuin field. Both direct and indirect
measurements of sirtuin activity are presented.
Finally, the
last five chapters (Part IV) on “Methods to study sirtuin biology” describe
protocols to measure some of the major biological pathways controlled by the sirtuins,
including metabolism, autophagy, genomic stability, circadian rhythms, and
calorie restriction.
Together, the
chapters in this volume present detailed protocols for sirtuin research that
can be followed directly or modified to investigate new areas of sirtuin
biology. We deeply thank the contributing authors to this volume and wish
success to the scientists using these protocols for discoveries in this rapidly
evolving and exciting field.
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