Figure 5. DNA-Bound Hsf1 Is Both Necessary and Sufficient to Drive Coalescence of a Pol II Gene.

(A) Left: physical map of HSP12 depicting the HSE and STREs within its upstream region (DNA motifs from www.yeastract.com); location of the deletion in hsp12-ΔHSE is indicated. Coordinates are numbered with respect to ATG. Right: Hsf1 ChIP-seq profile of the indicated region on Chr. VI in NHS and 5 min HS cells.

(B) Hsf1 ChIP analysis was conducted on HSP12 and hsp12-ΔHSE cells (strains AJ303 and AJ305, respectively) maintained at 30°C (NHS) or subjected to a 10-min HS as indicated. Depicted are means + SD (n = 2; qPCR = 4). *p < 0.05.

(C) Rpb1 ChIP analysis conducted as in (B). n.s., p > 0.05.

(D) Intragenic contact frequencies of HSP12 and hsp12-ΔHSE following 10 min HS as deduced by TaqI-3C (n = 2; qPCR = 4). n.s., p > 0.05.

(E) Intergenic contact frequencies between HSP12 or hsp12-ΔHSE and the indicated gene loci following 10 min HS. **p < 0.01.

(F) TaqI-3C analysis of HSP12 in Hsf1-AA cells pre-treated with 1 μM rapamycin for 90 min or not (+Rap or −Rap, respectively) followed by 10 min HS. Depicted are normalized frequencies of representative intragenic and intergenic interactions. *p < 0.05; n.s., p > 0.05.

(G) Physical map of the chromosomal transgene UAS_HS-BUD3.

(H) Hsf1 occupancy of BUD3 and UAS_HS-BUD3 (strains BY4741 and ASK804, respectively) under NHS and 10 min HS states. Analysis as in (B). **p < 0.05.

(I) Summary of intragenic interactions detected within UAS_HS-BUD3 in NHS and 10 min HS states (n = 2; qPCR = 4).

(J) Intergenic contact frequencies between BUD3 or UAS_HS-BUD3 and the indicated HSP genes in 10 min HS cells (n = 2; qPCR = 4).

See also Figures S1, S4, and S5.