Table 1.
Yeast strains and plasmids used
| Designation | Description |
|---|---|
| Strains | |
| NOY577 | MATα ade2 ade3 leu2 ura3 trp1 his can1 rrn5Δ::TRP1, pNOY103 (URA3, GAL7-35S rDNA) (4) |
| NOY798 | NOY577 carrying pNOY402 [LEU2, RRN5-(HA1)3-(His)6] instead of pNOY103 |
| NOY700 | MATa ade2-1 ura3-1 his3-11 trp1-1 leu2-3, 112 can1-100 rrn5Δ::LEU2, pNOY103; same as NOY699 (4) except for mating type |
| NOY844 | NOY700 carrying pNOY434 [HIS3, RRN5-(HA1)3-(His)6] instead of pNOY103 |
| MX1-4C | MATα ura3-52 leu2-3, 112 trp1 his3 Δ (hht1-hhf1) Δ(hht2-hhf2), pMS329 (CEN4 URA3 SUP11 HHT1-HHF1) (18) |
| NOY846 | MATα ura3 his3 trp1 leu2-3,112 rrn5Δ::LEU2Δ(hht1-hhf1) Δ(hht2-hhf2), pNOY434 [HIS3 RRN5-(HA1)3-(His)6], pMS329 (URA3 SUP11 HHT1-HHF1) |
| NOY847 | NOY846 carrying pNOY436 [TRP1 HHT2, myc-HHF2) instead of pMS329 |
| Plasmids | |
| pNOY103 | High-copy number plasmid carrying GAL7-35S rDNA, ADE3, URA3, 2μ, (2) |
| pNOY330 | LEU2, CEN6, RRN5-(HA1)3 (4) |
| pNOY402 | LEU2, CEN6, RRN5-(HA1)3-(His)6 derivative of pNOY330 in which an oligonucleotide encoding His tag (GSSHHHHHHSSG) is inserted immediately after the third HA1 repeat |
| pRS313 | Yeast-E. coli shuttle vector (19); HIS3, CEN6, ARS4 |
| pNOY434 | HIS3, CEN6, ARS4, RRN5-(HA1)3-(His)6, the 1.7-kb SalI/SacI fragment from pNOY402 carrying the His-tagged, HA1-tagged RRN5 gene cloned between the XhoI and SacI sites of pRS313 |
| pRS314 | Yeast-E. coli shuttle vector (19); TRP1, CEN6, ARS4 |
| pCC66 | Plasmid clone carrying the HHT2-HHF2 locus (a gift from F. Winston, Harvard; ref. 20) |
| pNOY435 | A 2.1-kb SacI/EcoRI fragment carrying HHT2 and HHF2 was amplified by PCR and cloned between the SacI and EcoRI sites of pRS314 |
| pNOY436 | TRP1, CEN6, ARS4, HHT2, myc-HHF2; oligonucleotide encoding a myc tag sequence (SEQKLISEEDL) inserted between the first and second codons of HHF2 in pNOY435 by site-directed mutagenesis |