TABLE 1

Proteins isolated in the targeted silencing screens

ProteinFused at/total residuesaCommentsReference
Aeb::G screen results
Sir1163/678Silencing proteinStone et al. (1991)
Rad794/566Binds Sir3Paetkau et al. (1994)
Rif11589/1917Binds Rap1Hardy et al. (1992)
Rif11742/1917Binds Rap1
Ris11398/1601Previously called Dis1; binds Sir4Zhang and Buchman (1997)
Ubp1078/791Also called Dot4; binds Sir4Singer et al. (1998)
Vac81/578Vacuolar protein
Esc11395/1658Binds Sir4Andrulis et al. (2002)
Esc2115/456Binds Sir2; SUMO-like domainCuperus and Shore (2002)
S-rich peptideb18/50
aeb::G screen results
Sir1163/678Silencing proteinStone et al. (1991)
Orc137/914Binds Sir1Triolo and Sternglanz (1996)
Rap165/827Binds Sir3 and Sir4Moretti et al. (1994); Moretti and Shore (2001)
Rap1637/827Binds Sir3 and Sir4
Rif11742/1917Binds Rap1Hardy et al. (1992)
Ris1398/1601Binds Sir4Zhang et al. (2002)
Ris1467/1601Binds Sir4
Hst145/503Sir2 homologBrachmann et al. (1995); Derbyshire et al. (1996)
Rpb487/221RNA polII subunit
Hir1475/840Histone gene repressorSherwood et al. (1993)
Esc2115/456Binds Sir2Cuperus and Shore (2002)
Esc4–1/1070Has six BRCT motifs
Net1566/1189Binds Sir2Cuperus and Shore (2002)
S-rich peptideb18/50
S-rich peptideb22/50
PeptidecIFLRLVKRPWPGQNFAP

Screens were performed as follows: Strains YEA76 (Aeb::G) or YEA78 (aeb::G) were transformed with a GBD yeast genomic library constructed in a derivative of the vector pGBT9c (Bartel et al. 1996). Trp+ transformants were selected and then replica plated to 5-fluoroorotic acid (5-FOA)-containing medium. 5-FOAR colonies were selected and assessed for plasmid dependence. Plasmid DNA was isolated from appropriate candidates and retransformed into the same yeast strain. Positive candidates were sequenced. Rif1 (fused at residue 1589) was identified six times, Ubp10 two times, Esc2 eight times, and the rest of the proteins one time.

  • a The number on the left indicates the amino acid residue in the protein at its junction point with GBD. The number on the right indicates the total number of amino acids in the protein. Each hybrid included all amino acids from the junction point to the C terminus of the protein.

  • b This peptide is encoded by the noncoding strand of the RSE1 gene, ORF YML049c. The sequence of this peptide beginning with residue 18 of a hypothetical (and most likely not naturally expressed) ORF is DICNISISSSISSSSSSLLSSPPPFSCLLTLDS.

  • c This peptide, whose sequence is shown, is not encoded by an ORF. The DNA coding for it is from near the 3′ end of the RGM1 gene.