Table 2.
Analysis of TBP–DNA complexes with various transcription factors
TBP mutant | Complex formationa | |||||
---|---|---|---|---|---|---|
TBP–DNA | BTAF1–TBP–DNA | BTAF1–TBP–mDNAb | TFIIA–TBP–DNA | NC2–TBP–DNA | TFIIB–TBP–DNA | |
wild-type | + | + | + | + | + | + |
V162A | − | + | + | + | ± | − |
L185A | − | + | − | ± | − | ± |
R186E | + | ± | − | + | ± | − |
R188E | + | − | − | + | − | + |
N189E | ± | − | n.d. | − | ± | + |
E191K | − | + | + | + | + | − |
K195A | + | + | n.d. | + | + | + |
R208E | − | + | n.d. | ± | + | − |
T210K | − | + | + | − | + | − |
V240A | ± | + | + | + | ± | ± |
Q242A | + | + | + | + | ± | ± |
K243E | + | ± | n.d. | − | + | + |
K249A | − | ± | − | − | − | − |
K254A | − | + | + | + | + | − |
K265A | − | + | + | + | + | ± |
F288K | − | ± | − | − | − | − |
R299A | − | + | + | + | + | ± |
R318A | − | + | + | + | + | − |
aAnalysed in EMSA as described in Materials and Methods; plus, >50% of the wild-type TBP–DNA complex with indicated factor; plus-minus, 25–50% of the wild-type TBP–DNA complex with indicated factor; minus, <25% of the wild-type TBP–DNA complex with indicated factor.
bmDNA—EMSA probe bearing the mutated TATA-box (CGCAAACG).
n.d., not determined.