Figure 5. HMS2 sense transcription represses the HMS2 antisense transcript and BAT2.

(A) Schematic showing transcripts when expression of HMS2 is regulated by the GAL1 promoter (pGAL1) in glucose (antisense-dominant state) or galactose (sense-dominant state) or when the HMS2 coding region is substituted with the URA3 coding region. (B, C, E, G–I) Northern blots showing total RNA (C, E, G, I) or poly(A)⁺ RNA (B, H) from cells expressing pGAL1:HMS2 cultured in glucose (GLU) or induced for the times indicated in galactose (GAL) in the genetic backgrounds indicated. (D, F) Quantitation of the experiments exemplified in (C) and (E) respectively, n = 2, error are SEM, Figure 5—source data 1A,B,C. (E, F) Anchor Away of Gal4 is achieved after incubation with rapamycin in DMSO for 1 h, added 3 h after induction with galactose (240 Rap). The control culture (240 DMSO) was treated with DMSO. See also Figure 5—figure supplement 1.

DOI: http://dx.doi.org/10.7554/eLife.03635.017

Figure 5—figure supplement 1. Mapping the 3′ ends of the pGAL.HMS2 antisense transcript isoforms.

(A) Schematic of the pGAL1.HMS2 transcription unit. (B) 3′RACE performed on the galactose-inducible HMS2 strain in glucose displays evidence of two poly(A) tails, one consistent with the short, B^G2, AS isoform (poly(A) tail²) terminating at a region downstream of the UAS_GAL (underlined in orange) ≈195 nt upstream of the GAL1 ATG and the long isoform, B^G1, terminating at a region upstream of the UAS_GAL (poly(A) tail¹), ≈500 nt upstream of the GAL1 ATG. (C) Evidence for an intermediate species, B^G3, not resolvable using Northern analysis, was detected after 3′ RACE was performed on cells grown in galactose for 3 hr. Potential end sites are marked with an asterisk.

DOI: http://dx.doi.org/10.7554/eLife.03635.019