Table 2.

Transcription analyses of the TAF25 mutant familya

StrainTotal poly (A)+ mRNASAGATFIIDTAF25p
YHB1PHO84YJL012CYDL124WPMT4WSC2TRX1GIC2YOR248WFAS2
Wild type124123394101233829044925296
G101E 22 22 24087160 37 36 5486 26 37
TAF25ΔN872633447168406445823431
TAF25ΔC984242264136557245636337
TAF25ΔN+ΔC8062324612358577565910757
86A77372287115350433676 26 55
88A 35 53 89 51285 41 30 5075 33 56
89A7945 166 5910262384253 38 40
90A78 28 3156026564384878 37 37
97A70523587027756 32 31545764
101A5250 139 56331 26 2 4852 24 32
106A7136 166 71 97 36 31 4646 37 29
108A81442705922446604747 45 28
118A78392056412152603555 46 35
119A63432205026745 25 4035 32 32
123A52 32 28656 135 41 26 47 17 38 37
127A62593636619748744163 35 26
184A83562366816162434059 41 32
188A54 21 34267 60 48 3 22 24 9 12
194A8156 124 6827565534652 34 36
195A59 21 435 51 97 24 27 27 48 30 27
196A72 30 28177145 34 34 32635749
202A834323561124 38 674155 43 34
  • a All analyses were performed three times using three independent cultures of the indicated yeast strains. RNA was either directly blotted and probed for total poly(A)+mRNA by hybridization with 32P-labeled oligo(dT)20 or fractionated by denaturing agarose gel electrophoresis, blotted, and detected with the indicated32P-labeled, PCR-generated gene-specific hybridization probes as detailed in Materials and Methods. TAF25ΔN+ΔC was the relevant control strain. Values that were at least 50% increased or 2-fold decreased relative to those for this control strain are in bold type; see the text for details. The values shown represent the means for three independent analyses; standard errors were <5% the indicated values for the total poly(A)+ mRNA analyses or <20% the indicated values for the RNA blotting experiments. The three classes of genes used for the transcription analyses were chosen based upon the data of Lee et al. (43). For TFIID-dependent genes PMT4, WSC2, andTRX1, transcription was reduced in a temperative-sensitivetaf145(130) strain after 45 min at a nonpermissive temperature 3.2-, 4.8-, and 5.3- fold (mRNA half-lives of 10, 11, and 18 min), respectively. For the SAGA-dependent genes YHB1, PHO84, YDL124W, and YJL012C, transcription was reduced in a gcn5 null strain 4.1-, 20.1-, 3.5-, and 3.0- fold (half-lives of 6, 7, 14, and 14 min), respectively. For the TAF25-dependent genes GIC2, YOR248W, and FAS2 transcription was reduced 4.2-, 4.1-, and 3.1- fold (half-lives of 36, 13 and 13).