Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD+ salvage pathway

Joseph J. Sandmeier, Ivana Celic, Jef D. Boeke, Jeffrey S. Smith

Research output: Contribution to journalArticle

Abstract

The Sir2 protein is an NAD+-dependent protein deacetylase that is required for silencing at the silent mating-type loci, telomeres, and the ribosomal DNA (rDNA). Mutations in the NAD+ salvage gene NPT1 weaken all three forms of silencing and also cause a reduction in the intracellular NAD+ level. We now show that mutation of a highly conserved histidine residue in Npt1p results in a silencing defect, indicating that Npt1p enzymatic activity is required for silencing. Deletion of another NAD+ salvage pathway gene called PNC1 caused a less severe silencing defect and did not significantly reduce the intracellular NAD+ concentration. However, silencing in the absence of PNC1 was completely dependent on the import of nicotinic acid from the growth medium. Deletion of a gene in the de novo NAD+ synthesis pathway BNA1 resulted in a significant rDNA silencing defect only on medium deficient in nicotinic acid, an NAD+ precursor. By immunofluorescence microscopy, Myc-tagged Bna1p was localized throughout the whole cell in an asynchronously growing population. In contrast, Myc-tagged Npt1p was highly concentrated in the nucleus in ̃40% of the cells, indicating that NAD+ salvage occurs in the nucleus in a significant fraction of cells. We propose a model in which two components of the NAD+ salvage pathway, Pnc1p and Npt1p, function together in recycling the nuclear nicotinamide generated by Sir2p deacetylase activity back into NAD+.

Original languageEnglish (US)
Pages (from-to)877-889
Number of pages13
JournalGenetics
Volume160
Issue number3
StatePublished - 2002

Fingerprint

Ribosomal DNA
NAD
Saccharomyces cerevisiae
Niacin
Mutation
Niacinamide
Telomere
Gene Deletion
Recycling
Fluorescence Microscopy
Histidine
Genes
Proteins
Growth

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Sandmeier, J. J., Celic, I., Boeke, J. D., & Smith, J. S. (2002). Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD+ salvage pathway. Genetics, 160(3), 877-889.

Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD+ salvage pathway. / Sandmeier, Joseph J.; Celic, Ivana; Boeke, Jef D.; Smith, Jeffrey S.

In: Genetics, Vol. 160, No. 3, 2002, p. 877-889.

Research output: Contribution to journalArticle

Sandmeier, JJ, Celic, I, Boeke, JD & Smith, JS 2002, 'Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD+ salvage pathway', Genetics, vol. 160, no. 3, pp. 877-889.
Sandmeier, Joseph J. ; Celic, Ivana ; Boeke, Jef D. ; Smith, Jeffrey S. / Telomeric and rDNA silencing in Saccharomyces cerevisiae are dependent on a nuclear NAD+ salvage pathway. In: Genetics. 2002 ; Vol. 160, No. 3. pp. 877-889.
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