Yêu Cầu Của NAD và SIR2 cho Việc Kéo Dài Tuổi Thọ Bằng Hạn Chế Calo ở Saccharomyces cerevisiae

10.1038/15458

10.1016/S0092-8674(00)80493-6

10.1101/gad.13.19.2570

10.1016/S0092-8674(00)81160-5

10.1038/35001622

10.1073/pnas.110148297

10.1073/pnas.97.12.6658

10.1128/MMBR.63.3.554-569.1999

10.1128/jb.154.2.1002-1004.1983

10.1016/0047-6374(80)90028-7

10.1016/0168-9525(91)90018-L

10.1046/j.1365-2958.1999.01538.x

For supplemental data see Science Online (www.sciencemag.org/feature/data/1052473.shl).

10.1093/emboj/17.12.3326

10.1093/emboj/17.7.1996

10.1093/emboj/17.13.3556

10.1016/S1097-2765(00)80472-4

10.1016/S0076-6879(99)08004-0

10.1021/bi9720134

10.1093/nar/24.13.2519

S.-J. Lin P.-A. Defossez L. Guarente data not shown.

10.1126/science.273.5271.59

B. P. Yu Modulation of Aging Processes by Dietary Restriction (CRC Press Boca Raton FL 1994).

10.1111/j.1532-5415.1999.tb03851.x

10.1126/science.285.5432.1390

All life-span analyses in this study were carried out on YPD plates with 2% glucose (unless otherwise stated) at least twice independently with more than 45 cells for each strain each time. Representative results are shown.

Strain PSY316 MAT α ura3-52 leu2-3 112 his3-Δ200 ade2-101 lys2-801 RDN1::ADE2 was made by P. Park (Guarente laboratory strain). W303AR MAT a ura3-1 leu2-3 112 his3-11 15 trp1-1 ade2-1 RDN1::ADE2 can1-100 has been described in (3). A364A MAT a ade1 ade2 ura1 his7 lys2 tyr1 gal1 and A364A cdc35-1 (BR214-4a) were obtained from the Yeast Genetic Stock Center (University of California Berkeley). SGY446 MAT a his3 leu2-3 112 ura3-52 trp1 ade8 tpk1 Δ ::ADE8 tpk2-63 (ts) tpk3::TRP1 and SGY446 TPK2 (SGY559) have been described in (16). Owing to the slow growth of cdc35-1 and tpk1 Δ tpk2-63 tpk3 Δ mutants at 30°C life-span analyses of A364A and SGY strains were carried out at 25°C. Strains bearing the cdc25-10 temperature-sensitive mutation were made as described (13).

All gene deletions in this study were done by replacing the wild-type genes with the kan r marker as described (20) and were verified by PCR using oligonucleotides flanking the genes of interest.

Ribosomal DNA recombination and rDNA circle assays were carried out at 30°C the temperature at which life-span assays were performed. At 25°C we did notice an increase in the life-span of cdc25-10 cells without a detectable decrease in recombination (21) consistent with previous findings that life-span is a more sensitive assay than recombination for Sir2p function (3).

We thank D. McNabb S.-I. Imai and members of the Guarente laboratory for suggestions; S. Garrett and K. Tatchell for plasmids and strains; and D. McNabb F. B. Johnson and B. Jegalian for critical reading of the manuscript. Supported by NIH the Ellison Medical Foundation the Seaver Institute and the Howard and Linda Stern Fund (L.G.); individual National Research Service Award 5-F32-AG05857-02 (S.-J.L.); and a Merck/MIT postdoctoral fellowship (P.-A.D.). P.-A.D. thanks G. Adelmant for continued support.