title: Pnc1p-mediated nicotinamide clearance modifies the epigenetic properties of rDNA silencing in Saccharomyces cerevisiae.
abstract: The histone deacetylase activity of Sir2p is dependent on NAD(+) and inhibited by nicotinamide (NAM). As a result, Sir2p-regulated processes in Saccharomyces cerevisiae such as silencing and replicative aging are susceptible to alterations in cellular NAD(+) and NAM levels. We have determined that high concentrations of NAM in the growth medium elevate the intracellular NAD(+) concentration through a mechanism that is partially dependent on NPT1, an important gene in the Preiss-Handler NAD(+) salvage pathway. Overexpression of the nicotinamidase, Pnc1p, prevents inhibition of Sir2p by the excess NAM while maintaining the elevated NAD(+) concentration. This growth condition alters the epigenetics of rDNA silencing, such that repression of a URA3 reporter gene located at the rDNA induces growth on media that either lacks uracil or contains 5-fluoroorotic acid (5-FOA), an unusual dual phenotype that is reminiscent of telomeric silencing (TPE) of URA3. Despite the similarities to TPE, the modified rDNA silencing phenotype does not require the SIR complex. Instead, it retains key characteristics of typical rDNA silencing, including RENT and Pol I dependence, as well as a requirement for the Preiss-Handler NAD(+) salvage pathway. Exogenous nicotinamide can therefore have negative or positive impacts on rDNA silencing, depending on the PNC1 expression level.
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, Virginia 22908, USA.
authors.forename: Julie M
authors.initials: JM
authors.lastname: McClure
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Gallo
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Smith
authors.forename: Mirela
authors.initials: M
authors.lastname: Matecic
authors.forename: Robert D
authors.initials: RD
authors.lastname: Hontz
authors.forename: Stephen W
authors.initials: SW
authors.lastname: Buck
authors.forename: Frances G
authors.initials: FG
authors.lastname: Racette
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Smith
mesh_terms: D004275:DNA, Ribosomal / Q000235:genetics* / Q000378:metabolism; D020868:Gene Silencing; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009536:Niacinamide / Q000378:metabolism*; D009535:Nicotinamidase / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D056565:Sirtuin 2; D037761:Sirtuins / Q000235:genetics / Q000378:metabolism; D016615:Telomere / Q000378:metabolism
chemical_list: D004275:DNA, Ribosomal; D029701:Saccharomyces cerevisiae Proteins; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae; D009243:NAD; D009536:Niacinamide; C087778:SIR2 protein, S cerevisiae; D056565:Sirtuin 2; D037761:Sirtuins; D009535:Nicotinamidase; C474330:PNC1 protein, S cerevisiae; D006655:Histone Deacetylases
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