Numbers indicate units of β-galactosidase activity. Those in bold face represent values obtained in the original screens; others were derived from the subsequent systematic analyses. For the DBD-septin fusions, F denotes the full-length septins, N denotes fragments extending from the N-terminus to the N-terminal boundary of the predicted coiled-coil domains, and C denotes C-terminal fragments consisting essentially of the predicted coiled-coil domains. A full-length AD-anillin clone (see Materials and Methods) and the empty vector pJG4-5 were also used as negative controls with each of the DBD fusions; in all cases, these yielded low values similar to those obtained with the full-length DmUba2.

10 positive clones from the screens proved to contain fragments of pnut. The two isolated with Pnut as bait contained sequences beginning at amino acids 78 and 142 but were not completely sequenced. All of the eight isolated with Sep2 as bait contained sequences beginning between amino acids 413 and 431 (of the 539-amino-acid protein) but were not completely sequenced.

Seven positive clones from the screens proved to contain fragments of sep2. All of the six isolated with Pnut as bait contained sequences beginning between amino acids 265 and 288. In at least two of these, the sequences appeared to run to the C-terminus; the others were not completely sequenced. The clone isolated using Sep1 as bait contained sequences starting at amino acid 27 but was not completely sequenced.

Three positive clones from the screens proved to contain fragments of Dmuba2. Two of these (one from the Sep1 screen and one from the Sep2 screen; interaction values shown in the table) encoded amino acids 552-700;one of these clones was used in the systematic two-hybrid analyses. The third Dmuba2 clone (from the Sep1 screen; 708 units of β-galactosidase activity) encoded amino acids 427-700.

Full-length AD-DmUba2 (see Materials and Methods andTable 3).

The single Dmubc9 clone isolated in the screen contained the complete ORF (see text).

Each of these interactors was represented by a single positive clone from the screen.

Three positive clones from the Sep1 screen contained identical fragments of the gene designated sip4. The original interaction value for one of these clones is shown. This same AD-fusion clone was used in the systematic two-hybrid analyses.