TABLE 4

Effect of ime1 mutations of Ime1p-Ume6p interaction

GBD-fusionaTwo-hybrid interaction with GAD-Ume6pb (β-galactosidase activityc)
None0.77
Ime1p422
Ime1-L321Fp29
Ime1-R347Kp1.5
Ime1-Q340*p2.8
Ime1-L321F, S360Fp221
Ime1-R347K, S360Fp42
Ime1-S352A, S356A, S360Ap214
Ime1-ala8p0.4
  • a GBD-Ime1p includes residues 294–360 from Ime1p. GBD-Ime1p derivatives yielded background β-galactosidase levels of 0.2 to 1.2, as determined with GAD lacking Ume6p residues.

  • b GAD-Ume6p includes residues 1–161 from Ume6p.

  • c Strain Y190 was used in these assays. β-Galactosidase activity was determined in log phase cultures in YPAc medium. The values, in Miller units, are the mean of three determinations and had standard deviations of <30% (for values <200 units) or <20% (for values >200 units).