An Uncharacterized Domain Within the N-Terminal Tail of Histone H3 Regulates the Transcription of FLO1 via Cyc8

Yeast flocculation relies on cell surface flocculin proteins encoded by the sub-telomeric gene, FLO1. The expression of FLO1 is antagonistically regulated by the Tup1-Cyc8 repressor complex and the Swi-Snf co-activator complexes. The role of hyperacetylated N-terminal amino acid residues of histone H3 and H4 is well established in the transcription of FLO1 and other Tup1-Cyc8 regulated genes. However, sub-domains within the tails of histone H3 and H4 are yet to be identified and the mechanism by which they regulate the FLO1 transcription is completely unexplored. Upon screening of different H3 and H4 N-terminal stretch deletion mutants, we have identified a new region within the N-terminal tail of histone H3, H3 Delta(17-24) regulating the transcription of FLO1 and FLO5. This N-terminal truncation mutant showed higher FLO1 and FLO5 expression by 68% and 41% respectively compared to wild-type H3. Further examination showed reduced Cyc8 and nucleosome occupancy in the upstream regulatory region of active flo1 in the H3 Delta(17-24) mutant than in H3 wild-type cells. The findings also indicate that Hda1 assists in Cyc8 interaction at the active FLO1 template. Altogether we demonstrate that Tup1-independent interaction of Cyc8 with the active FLO1 gene acts as a transcription limiting factor and that the histone H3 N-terminal 17-24 stretch is essential for this interaction. In the absence of the 17-24 stretch, the Cyc8 restrictive effect is altered, resulting in over-expression of FLO1.