Table 2. Methyltransferase inhibits ORF3 binding to the X-domain and vice versa.
| Yeast cotransformants | LT− + 1mM M | LTHA−+1mM M | LTH−Ar++1mM M | LT−+X-α-gal +1mM M | LTH− + 1mM M + 3-AT (mM) |
LTM− | LTHAM− | LTHM−Ar+ | LTM−+X-α-gal | LTHM− + 3-AT (mM) |
||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 5 | 10 | 5 | 10 | |||||||||
| BD ORF3 Met + AD X | +++ | +++ | +++ | +++ | +++ | +++ | +++ | − | − | − | − | − |
| BD ORF3 + AD X | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ | +++ |
| BD Met ORF3 + AD X | +++ | ++ | + | + | ++ | − | +++ | − | − | − | − | − |
| BD Met + AD X | +++ | ++ | + | + | ++ | − | +++ | ++ | + | + | ++ | − |
| BD Met ORF3 + AD | +++ | − | − | − | − | − | +++ | − | − | − | − | − |
| BD Met + AD | +++ | − | − | − | − | − | +++ | − | − | − | − | − |
| BD ORF3 Met + AD | +++ | − | − | − | − | − | +++ | − | − | − | − | − |
| BD ORF3 + AD | +++ | − | − | − | − | − | +++ | − | − | − | − | − |
| BD Met + AD ORF3 | +++ | − | − | − | − | − | +++ | − | − | − | − | − |
Y2H gold strain was transformed in indicated combinations and plated on media lacking leucine, tryptophan (LT−) and supplemented with 1mM methionine (M). Eight random colonies from each cotransformants were replica plated onto media containing various selection markers, as indicated and their growth monitored over a period of four days. Cotransformants were grown on indicated media in the presence of 1mM methionine (M) or without methionine (M−). AD: Activation domain, BD: Binding domain, +++: Strong growth, ++: Moderate growth, +: Poor growth, −: No growth, L: Leucine, T: Tryptophan, H: Histidine, A: Adenine hemisulfate, M: Methionine, Ar: Aureobasidin, 3-AT: 3-amino 1, 2, 4 Triazole, “−”: Deficiency in the medium, “+”: Supplemented in the medium.