title: Involvement of Daphnia pulicaria Sir2 in regulating stress response and lifespan.
abstract: The ability to appropriately respond to proteotoxic stimuli is a major determinant of longevity and involves induction of various heat shock response (HSR) genes, which are essential to cope with cellular and organismal insults throughout lifespan. The activity of NAD+-dependent deacetylase Sir2, originally discovered in yeast, is known to be essential for effective HSR and longevity. Our previous work on HSR inDaphnia pulicaria indicated a drastic reduction of the HSR in older organisms. In this report we investigate the role of Sir2 in regulating HSR during the lifespan of D. pulicaria. We cloned Daphnia Sir2 open reading frame (ORF) to characterize the enzyme activity and confirmed that the overall function of Sir2 was conserved in Daphnia. The Sir2 mRNA levels increased while the enzyme activity declined with age and considering that Sir2 activity regulates HSR, this explains the previously observed age-dependent decline in HSR. Finally, we tested the effect of Sir2 knockdown throughout adult life by using our new RNA interference (RNAi) method by feeding. Sir2 knockdown severely reduced both the median lifespan as well as significantly increased mortality following heat shock. Our study provides the first characterization and functional study of Daphnia Sir2.
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA.
authors.forename: Charles A
authors.initials: CA
authors.lastname: Schumpert
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Anderson
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA.
authors.forename: Jeffry L
authors.initials: JL
authors.lastname: Dudycha
authors.affiliation: Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA.
authors.forename: Rekha C
authors.initials: RC
authors.lastname: Patel
mesh_terms: D000375:Aging / Q000502:physiology*; D000818:Animals; D015153:Blotting, Western; D003621:Daphnia / Q000502:physiology*; D055785:Gene Knockdown Techniques; D018869:Heat-Shock Response / Q000502:physiology*; D008136:Longevity / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D056565:Sirtuin 2 / Q000378:metabolism*
chemical_list: D056565:Sirtuin 2
keywords: Daphnia; RNAi; Sir2; heat shock; lifespan