Table 2.

Plasmids used in this study

PlasmidsCharacteristics (reference)
pRS314 TRP1 CEN6 (60)
pRS316 URA3 CEN6 (60)
pKO11 PGAL1-HA2-TTDH3 URA3 CEN6 (29)
pGGX1 PGAL1-EGFP-TTDH3 URA3 CEN6; made by replacing the 90-bpEcoRI-BamHI fragment containing two HA epitopes of pK011 with the 0.72-kb EcoRI-BamHI fragment containing the enhanced GFP coding region without the termination codon, which was amplified by PCR using pEGFP-N1 (Clontech) as a template
pAGX1 PACT1-EGFP-TTDH3URA3 CEN6; made by replacing the 720-bpNotI-EcoRI fragment containingPGAL1 of pGGX1 by the 0.47-kbNotI-EcoRI PCR fragment containingPACT1
pAGX2 PACT1-EGFP-(A4GA4G)-TTDH3URA3 CEN6; made by inserting the BglII-BamHI synthetic double-stragonucleotide of 5′-AGATCTGCTGCCGCTGCTGGTACCGCTGCTGCCGCTGGATCC-3′ (each underlined region corresponds to the recognition site ofBglII, KpnI, and BamHI, respectively), which encodes the linkerpolypeptide of RSAAAAGTAAAAGS at theBamHI site of pAGX1
pTAGX2 PACT1-EGFP-(A4GA4G)-TTDH3 TRP1 CEN6; made by replacing the NotI-XhoI fragment of the multicloning site of pRS314 by theNotI-XhoI fragment containingPACT1-EGFP-(A4GA4G)-TTDH3 of pAGX2
pAGX2-BNI1 PACT1-EGFP-(A4GA4G)-BNI1-TTDH3URA3 CEN6; made by inserting the 5.9-kbBamHI-SmaI fragment from pRS314-PBNI1-myc-BNI1 containing the BNI1 open reading frame into the corresponding sites of pAGX2
pTAGX2-BNI1 PACT1-EGFP-(A4GA4G)-BNI1-TTDH3TRP1 CEN6; made by inserting the 5.9-kbBamHI-SmaI fragment from pRS314-PBNI1-myc-BNI1 containing the BNI1 open reading frame into the corresponding sites of pTAGX2
pGGX1-BNI1 PGAL1-EGFP-BNI1-TTDH3URA3 CEN6; made by inserting the 5.9-kbBamHI-SmaI fragment from pRS314-PBNI1-myc-BNI1 containing the BNI1 open reading frame into the corresponding sites of pGGX1
pAGX1-ABP1 PACT1-EGFP-ABP1-TTDH3URA3 CEN6; made by inserting the 1.8-kbSmaI-SmaI PCR fragment containing theABP1 open reading frame into the corresponding site of pAGX1
pFA6a-His3MX6 PTEF1-his5 +-TTEF1 (44)
pRS314-PBNI1-myc PBNI1-myc3-TTDH3TRP1 CEN6 (18)
pRS314-PBNI1-myc-BNI1 PBNI1-myc3-BNI1-TTDH3TRP1 CEN6 (18)
pRS314-PBNI1-myc-bni1 (490-1954) PBNI1-myc3-bni1 (490-1954)-TTDH3 TRP1 CEN6 (18)
pRS314-PBNI1-myc-bni1 (Δ133-245) PBNI1-myc3-bni1133-245)-TTDH3 TRP1 CEN6; made by deleting the 339-bp BsaHI-BsaHI fragment from pRS314-PBNI1-myc-BNI1
pRS314-PBNI1-myc-bni1 (Δ826-987) PBNI1-myc3-bni1 (Δ826-987)-TTDH3 TRP1 CEN6 (18)
pRS314-PBNI1-myc-bni1 (Δ1239-1328) PBNI1-myc3-bni1 (Δ1239-1328)-TTDH3 TRP1 CEN6 (18)
pRS314-PBNI1-myc-bni1 (1-1750) PBNI1-myc3-bni1 (1-1750)-TTDH3 TRP1 CEN6 (18)
pUC19-BUD6 BUD6; made by inserting the 3.8-kbSalI-SmaI PCR fragment containing 0.74-kbBUD6 upstream and 0.66-kb downstream noncoding regions into the SalI-SacI (blunted) site of pUC19
pUC19-bud6::HIS3a derivative of pUC19-BUD6; made by replacing the 2.2-kb NheI-SacI fragment ofBUD6 with the 1.8-kb HIS3 fragment