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Nucleosome structure of the yeast CHA1 promoter: analysis of activation‐dependent chromatin remodeling of an RNA‐polymerase‐II‐transcribed gene in TBP and RNA pol II mutants defective in vivo in response to acidic activators
Author(s) -
Moreira José M.A.,
Holmberg Steen
Publication year - 1998
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/17.20.6028
Subject(s) - biology , rna polymerase ii , rna polymerase iii , rna polymerase i , chromatin , chromatin remodeling , nucleosome , microbiology and biotechnology , small nuclear rna , rna polymerase , rna , genetics , polymerase , gene , rna dependent rna polymerase , gene expression , promoter
The Saccharomyces cerevisiae CHA1 gene encodes the catabolic L‐serine (L‐threonine) dehydratase. We have previously shown that the transcriptional activator protein Cha4p mediates serine/threonine induction of CHA1 expression. We used accessibility to micrococcal nuclease and DNase I to determine the in vivo chromatin structure of the CHA1 chromosomal locus, both in the non‐induced state and upon induction. Upon activation, a precisely positioned nucleosome (nuc‐1) occluding the TATA box and the transcription start site is removed. A strain devoid of Cha4p showed no chromatin alteration under inducing conditions. Five yeast TBP mutants defective in different steps in activated transcription abolished CHA1 expression, but failed to affect induction‐dependent chromatin rearrangement of the promoter region. Progressive truncations of the RNA polymerase II C‐terminal domain caused a progressive reduction in CHA1 transcription, but no difference in chromatin remodeling. Analysis of swi1 , swi3 , snf5 and snf6 , as well as gcn5 , ada2 and ada3 mutants, suggested that neither the SWI/SNF complex nor the ADA/GCN5 complex is involved in efficient activation and/or remodeling of the CHA1 promoter. Interestingly, in a sir4 deletion strain, repression of CHA1 is partly lost and activator‐independent remodeling of nuc‐1 is observed. We propose a model for CHA1 activation based on promoter remodeling through interactions of Cha4p with chromatin components other than basal factors and associated proteins.