Yeast HAP1 activator binds to two upstream activation sites of different sequence

Pfeifer, Karl; Prezant, Toni R.; Guarente, Leonard

doi:10.1016/0092-8674(87)90751-3

Cited by 302 publications

(189 citation statements)

References 39 publications

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“…The hypersensitive site was detectable in a cells, in which nucleosomes were not positioned, but it was strongly diminished in α cells, indicating that Hap1 binding is severely limited at the dyad in the positioned nucleosome IV. Similar results for Hap1 binding were obtained for TALS-UAS1b (lanes [19][20][21][22][23][24] and -UAS1c (lanes [28][29][30][31][32][33], in which the UAS1 site was located in the peripheral region of the positioned nucleosome IV; that is, the hypersensitive site is present in a cells, but absent in α cells. It should be noted that the location of UAS1 in TALSUASc is shifted to 5 bp exterior to the nucleosome edge, as compared to that in TALS-UAS1b.…”

Section: Effect Of the Uas1 Locations In A Positioned Nucleosome On Hsupporting

confidence: 80%

“…These minichromosomes have been utilized to examine the binding of Gal4 [7][8][9][10][11] and Bicoid [10], as well as to determine the effect of DNA sequences on nucleosome formation [13] in vivo. We chose to examine the binding of the Hap1 activator [24], which is a member of a family of fungal transcription factors with the Zn 2 Cys 6 binuclear cluster domain [25]. The UAS1 site was introduced at locations 24-43 bp, 29-48 bp and 61-80 bp interior to the edge (1522 nt) of the positioned nucleosome IV in TALS-UAS1a, TALSUAS1b and TALS-UAS1c, respectively ( Fig.…”

Section: Experimental Designmentioning

confidence: 99%

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A nucleosome positioned by α2/Mcm1 prevents Hap1 activator binding in vivo

Morohashi

Nakajima

Kurihara

et al. 2007

Biochemical and Biophysical Research Communications

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Nucleosome positioning has been proposed as a mechanism of transcriptional repression. Here, we examined whether nucleosome positioning affects activator binding in living yeast cells. We introduced the cognate Hap1 binding site (UAS1) at a location 24-43 bp, 29-48 bp or 61-80 bp interior to the edge of a nucleosome positioned by α2/Mcm1 in yeast minichromosomes. Hap1 binding to the UAS1 was severely inhibited, not only at the pseudo-dyad but also in the peripheral region of the positioned nucleosome in α cells, while it was detectable in a cells, in which the nucleosomes were not positioned. Hap1 binding was restored in α cells with tup1 or isw2 mutations, which caused the loss of nucleosome positioning. These results support the mechanism in which α2/Mcm1-dependent nucleosome positioning has a regulatory function to limit the access of transcription factors.Keywords nucleosome positioning; chromatin; activator binding; in vivo footprinting; yeast In a general view of the process of transcriptional activation, activators have to gain access to an array of nucleosomes, and chromatin remodeling complexes and/or histone modification enzymes change the structural features of chromatin to recruit transcriptional machinery to the promoter regions [reviewed in 1,2]. Regarding activator binding, several regulatory factors such as steroid hormone receptor, Gal4-derivatives, USF and Sp1, bind to reconstituted nucleosomes in vitro under some circumstances, although their affinities for nucleosomal DNA decrease by one to three orders of magnitude, relative to those for naked DNA [see reviews, 2-4]. Positioned nucleosomes assembled with Drosophila embryo extracts preclude NF1 binding to its cognate site [5]. In contrast, NF-κB binds to its recognition sites within a positioned nucleosome with an affinity close to that for free DNA in vitro [6]. Yeast Gal4 is one of the best characterized activators binding to nucleosomes in vivo. Gal4 can bind to its cognate site within a positioned nucleosome to perturb the nucleosome in yeast cells [7][8][9][10][11]. Also, the Pho4 activator can reportedly bind its nucleosome-occupied site before nucleosome disassembly in vivo [12]. However, only a few activators have been examined for their access *Corresponding author. Mitsuhiro Shimizu, Department of Chemistry, Meisei University, 2-1-1 Hodokubo, Hino, Tokyo 191-8506, Japan, Phone: +81-42-591-7483, Fax: +81-42-591-7419, E-mail: shimizum@chem.meisei-u.ac.jp. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. to DNA within the nucleosome in vivo. Although nucleosome positioning has been observ...

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Section: Effect Of the Uas1 Locations In A Positioned Nucleosome On Hsupporting

confidence: 80%

Section: Experimental Designmentioning

confidence: 99%

A nucleosome positioned by α2/Mcm1 prevents Hap1 activator binding in vivo

Morohashi

Nakajima

Kurihara

et al. 2007

Biochemical and Biophysical Research Communications

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“…There are many precedents for the ability of a DNA-binding protein to recognize unrelated sequences (54)(55)(56). A possible scenario is that the regulatory effects of YB-1 may vary with the target sequence and YB-1 could serve as either a positive or a negative regulator of gene expression depending on the target sequence (57).…”

Section: And Class II Mhc Genes (33) Ifn-"(-induced Li Gene Transcmentioning

confidence: 99%

YB-1 DNA-binding protein represses interferon gamma activation of class II major histocompatibility complex genes.

Ting

Painter

Zeleznik‐Le

et al. 1994

The Journal of Experimental Medicine

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SummaryInterferon y (IFN-'r) is the most potent inducer of class II major histocompatibility complex (MHC) genes. This induction is uniquely mediated by three DNA elements in the promoter region of class II MHC genes. One of these DNA elements, Y, contains an inverted CCAAT box. Previously, we have screened a Xgt11 library for Y-binding proteins and identified the YB-1 gene. Here we provide evidence that YB-1 can repress the IFN-3~ induction of class II MHC promoter as well as the Invariant chain (Ii) gene which also contains a Y element in its promoter. This was demonstrated by cotransfecting a YB-1 expression vector with promoter-reporter gene constructs. As an alternate approach, an effcient transient transfection system was developed which resulted in a )70% transfection efficiency. Transfection of YB-1 by this procedure resulted in the near abrogation of IFN-7 induced HLA-DR antigen and mRNA expression. These findings show the functional suppression of class II MHC gene induction by the YB-1 protein.C lass II MHC gene products play a variety of important roles in immune regulation (reviewed in 1-3). These molecules control the acquisition of the mature T cell repertoire and serve as restriction elements for CD4 § T cells. These functions place the regulation of class II MHC antigens as an important topic in immune regulation. The expression of class II MHC genes is primarily regulated at the level of transcription. In the past few years, we and others have delineated an array of cis-acting elements important for optimal class II gene regulation and have identified proteins that bind to these elements. The cis-acting regulatory elements of the DRA gene are probably the best analyzed, and include three elements (S, X, and Y) that are also found in other class II MHC promoters (4-7). The X and Y elements constitute the conserved class II box present in the upstream region of all class II MHC promoters studied to date (8, 9). These two elements are separated by a 19-21-bp spacing that is conserved in length but not in sequence. The Y element contains an inverted CCAAT element, and the X element has been functionally divided into two subregions, X1 and X2, based on the separate interactions of these subregions with the RF-X and hXBP-1 recombinant DNA-binding proteins, respectively (10, 11). The S sequence (also known as H, or W/Z which is a larger DNA sequence) is a heptamer sequence located upstream of the class II box (12-17). These three elements are important for basal gene transcription, and thus far, they are inseparable from elements required for the IFN-T induction of class II MHC genes (12-19).The YB-1 DNA-binding protein was initially identified by using radiolabeled Y element sequence to screen a ~,gt11 expression cDNA library (20). The recombinant YB-1 protein exhibits specificity for the Y element because mutations of an inverted CCAAT sequence in the Y element can abrogate its ability to interact with YB-1. An intriguing feature is the inverse relationship between levels of YB-1 and DRA in IFN-3,-a...

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“…The mode of Hap1 binding to its activated genes is well understood [18][19][20][21]. The promoters of many Hap1-activated genes generally contain a direct repeat of two CGG triplets separated by a six nucleotide spacer (optimal site: CGGnnnTAnCGG).…”

Section: Introductionmentioning

confidence: 99%

Regulation of the HAP1 gene involves positive actions of histone deacetylases

Xin

Lan

Lee

et al. 2007

Biochemical and Biophysical Research Communications

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The yeast transcriptional regulator Hap1 promotes both transcriptional activation and repression. Previous studies have shown that Hap1 binds to the promoter of its own gene and represses its transcription. In this report, we identified the DNA site that allows Hap1 binding with high affinity. This Hap1-binding site contains only one CGG triplet and is distinct from the typical Hap1-binding upstream activation sequences (UASs) mediating transcriptional activation. Furthermore, at the HAP1 promoter, Ssa is bound to DNA with Hap1, whereas Hsp90 is not bound. Intriguingly, we found that histone deacetylases, including Rpd3, Hda1, Sin3 and Hos1, are not required for the repression of the HAP1 gene by Hap1. Rather, they are required for transcriptional activation of the HAP1 promoter, and this requirement is dependent on the HAP1 basal promoter. These results reveal a complex mechanism of transcriptional regulation at the HAP1 promoter, involving multiple DNA elements and regulatory proteins.

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Yeast HAP1 activator binds to two upstream activation sites of different sequence

Cited by 302 publications

References 39 publications

A nucleosome positioned by α2/Mcm1 prevents Hap1 activator binding in vivo

A nucleosome positioned by α2/Mcm1 prevents Hap1 activator binding in vivo

YB-1 DNA-binding protein represses interferon gamma activation of class II major histocompatibility complex genes.

Regulation of the HAP1 gene involves positive actions of histone deacetylases

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