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Vol. 13, No. 9, pp. 1089-1101, May 1, 1999
Department of Molecular Biology, Princeton University,
Princeton, New Jersey 08544 USA
The HM loci in Saccharomyces cerevisiae constitute
region-specific but gene-nonspecific repression domains, as a number of heterologous genes transcribed by RNA polymerase II or III are silenced
when placed at these loci. The promoters of the Ashbya gossypii
TEF gene and the S. cerevisiae TEF1 and TEF2 genes,
however, are resistant to transcriptional silencing by the HM
silencers in yeast. Moreover, when interposed between the HML
genes and the E silencer, certain segments of these
promoters block the repression effect of the silencer on the genes.
All of these fragments contain UASrpg (upstream activation
sequence of ribosome protein genes) composed of multiple binding sites
for Rap1. In fact, a 149-bp segment consisting essentially of only
three tandem Rap1-binding sites from the UASrpg of yeast
TEF2 exhibits silencer-blocking activity. This element also
exhibits insulating activity and orientation dependence characteristic
of known chromatin boundary elements. Finally, the element blocks the
physical spread of heterochromatin initiated at a silencer. This
segment provides the first example of chromatin domain boundary or
insulator elements in yeast.
[Key Words: Chromatin boundary elements; promoters; silencers; DNA topology; Rap1; transcriptional silencing]
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