TABLE 1.

Mapping of the IE2-Ubc9 interaction domain by yeast two-hybrid screening

Bait (pHybLex fusion)Prey (pYESTrp fusion)
Ubc9Ubc9 C93S
His-negative growthaβ-Gal activitybHis-negative growthaβ-Gal activityb
Control (empty vector)c1.00 ± 0.381.02 ± 0.17
IE2 1-720+35.6 ± 7.62*+38.8 ± 7.30*
IE2 925-1466+9.40 ± 2.89*+14.6 ± 2.31*
IE2 925-1233+7.73 ± 2.41*+10.0 ± 1.92*
IE2 925-1176+4.23 ± 1.48*+6.00 ± 1.19*
IE2 925-1131+4.09 ± 1.74*+6.81 ± 1.34*
IE2 925-1083+7.46 ± 2.25*+7.89 ± 1.64*
IE2 925-1037+14.4 ± 2.83*+20.9 ± 4.50*
IE2 925-9881.45 ± 0.511.52 ± 0.40
  • a A + indicates growth, while a − indicates that there was no growth.

  • b Results are expressed as means ± standard deviations from three distinct experiments. Reference activity was that of control cells cotransformed with pYESTrp-Ubc9 and pHybLex. Asterisks indicate that P was <0.05 compared with reference activity. β-Gal, β-galactosidase.

  • c Control cotransformations with empty prey vector pYESTrp and each of the bait constructs were all negative for His-negative growth except for pHybLex-IE2 1-720 (β-Gal activity = 41.0 ± 10.8; P < 0.05). Cotransformations with prey pYESTrp-SUMO-1 and each of the bait constructs were all negative for His-negative growth, except for pHybLex-IE2 1-720 (β-Gal activity = 39.5 ± 11.1; P < 0.05) and pHybLex-Ubc9 (12.1 ± 2.05; P < 0.05).