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. 2001 Apr 15;15(8):955–967. doi: 10.1101/gad.873601

Figure 3.

Figure 3

ORC was required for Sum1-1p-mediated silencing. (A) MATα haploids of the genotypes SIR SUM1 (W303) and sir2Δ SUM1-1 (JRY7152) were transformed with a plasmid vector (pRS316, line 1) or the vector containing HMRa (pJR759, line 2), HMRa-eΔ (pJR760, line 3), HMRa-syntetic silencer (pJR900, line 4), HMRa -no Rap1p/Abf1p binding sites (pJR891, line 5), or HMRa-synthteic silencer/no ORC binding site (pJR901, line 6). Representative transformants were tested for their mating ability by replica plating onto an a mating-type tester lawn (TD4) on minimal medium requiring plasmid retention for growth. Mating was indicated by growth of prototrophic diploids. (B) MATα haploids of the genotypes SIR SUM1-1 (MC89), sir2Δ SUM1-1 (JRY7152), sir3Δ SUM1-1 (JRY7161), sir4Δ SUM1-1 (JRY7166), SIR orc5-1 SUM1-1 (JRY7149), sir2Δ orc5-1 SUM1-1 (JRY7157), sir3Δ orc5-1 SUM1-1 (JRY7164), sir4Δ orc5-1 SUM1-1 (JRY7169), SIR orc2-1 SUM1-1 (JRY7150), sir2Δ orc2-1 SUM1-1 (JRY7159), orc5-1 (JRY7146), and orc2-1 (JRY7147) were tested for their mating ability as in A using mating tester JRY2726. (C) MATα haploids of the genotypes SIR SUM1 (W303), sir2Δ myc-SUM1-1 (JRY7153), sir2Δ myc-SUM1-1 orc1Δ1-235 (orc1ΔN, JRY7176), and sir2Δ myc-SUM1-1 ORC1 (Q236H, I237M) (orc1**, JRY7177) were tested for their mating ability as in B.