Abstract:
:DNA fragments containing the silencers that flank the mating type genes at HML alpha are shown to bind specifically to the nuclear scaffold of yeast. The scaffold proteins are solubilized with urea and then renatured to form a soluble extract which allows reconstitution of sequence-specific DNA loops. At the silent mating type locus HML alpha, loops are formed by either silencer-silencer (E-I) interaction or silencer-promoter interactions (E-P and I-P). The nuclear protein RAP-1 fractionates efficiently with the nuclear scaffold, and binds to the E, I, and promoter regions. Affinity purification of RAP-1 and oligonucleotide competition show that RAP-1 is necessary for reconstitution of loops in vitro. These results are consistent with a model in which silencers define a chromatin loop within which occur modifications that maintain the promoter in an inactive state.
journal_name
Celljournal_title
Cellauthors
Hofmann JF,Laroche T,Brand AH,Gasser SMdoi
10.1016/0092-8674(89)90788-5subject
Has Abstractpub_date
1989-06-02 00:00:00pages
725-37issue
5eissn
0092-8674issn
1097-4172pii
0092-8674(89)90788-5journal_volume
57pub_type
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