The C-terminal domain of SIN1 in yeast interacts with a protein that binds the URS1 region of the yeast HO gene

Yona, Eyal; Bangio, Haim; Erlich, Porat M.; Tepper, Steven H.; Katcoff, Don J.

doi:10.1007/bf00290726

Cited by 8 publications

(13 citation statements)

References 20 publications

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“…1). It was shown that one of these domains (amino acids 100 -162) was able to bind double-stranded DNA without substantial sequence specificity (6,11). Some key features of HMG boxes, however, were not present in this sequence, and its similarity to HMG was disputed (12).…”

mentioning

confidence: 99%

Functional Domains of the Yeast Chromatin Protein Sin1p/Spt2p Can Bind Four-way Junction and Crossing DNA Structures

Novoseler

Hershkovits

Katcoff

2005

Journal of Biological Chemistry

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Sin1p/Spt2p is a yeast chromatin protein that, when mutated or deleted, alters the transcription of a family of genes presumably by modulating local chromatin structure. In this study, we investigated the ability of different domains of this protein to bind four-way junction DNA (4WJDNA) since 4WJDNA can serve as a model for bent double helical DNA and for the crossed structure formed at the exit and entry of DNA to the nucleosomes. Sequence alignment of Sin1p/Spt2p homologues from 11 different yeast species showed conservation of several domains. We found that three domains of Sin1p/ Spt2p fused to glutathione S-transferase can each bind independently in a structure-specific manner to 4WJDNA as measured in a gel mobility shift assay. A feature common to these domains is a cluster of positively charged amino acids. Modification of this cluster resulted in either abolishment of binding or a change in the binding properties. One of the domains tested clearly bound superhelical DNA, although it failed to induce bending in a circularization assay. Poly-L-lysine, which may be viewed as a cluster of positively charged amino acids, bound 4WJDNA as well. Phenotypic analysis showed that disruption of any of these domains resulted in suppression of a his4-912␦ allele, indicating that each domain has functional significance. We propose that Sin1p/Spt2p is likely to modulate local chromatin structure by binding two strands of double-stranded DNA at their crossover point.Sin1p/Spt2p is a yeast chromatin non-histone protein. While the precise function of the protein is still unknown, it is known to function as a negative transcriptional regulator of a number of genes including SUC2 (1), INO1 (2), and SSA3 (3). Activity of the HO promoter, as measured from an HO promoter driving a lacZ gene (4), is also regulated by SIN1/SPT2. swi1, swi2, and swi3 mutants are unable to transcribe RNA from the HO promoter. However, in sin1/swi double mutants, transcription is restored. Mutants in SPT2 were first identified as suppressors of ty and ␦ insertions in the 5Ј non-coding region of the HIS4 gene (5). As the negative regulation of these genes is overcome by the SW1/SNF chromatin remodeling complex, it was suggested that a function of SIN1p/SPT2p is to somehow maintain chromatin compaction at specific locations in the chromatin.

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mentioning

confidence: 99%

Functional Domains of the Yeast Chromatin Protein Sin1p/Spt2p Can Bind Four-way Junction and Crossing DNA Structures

Novoseler

Hershkovits

Katcoff

2005

Journal of Biological Chemistry

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“…We have shown previously that the XBS can bind a protein that interacts with the C-terminal of SIN I [8]. A bacterially produced mutant sin 1-2 protein containing a single amino acid difference from the wild type SIN I was able to remove this protein from the XBS [4].…”

Section: Events At Tke )Ibs Cause the Overexpression Of Ho In Sin12 mentioning

confidence: 95%

“…HO) while it has not been shown to bind DNA in a sequencespecific manner. Previously we showed that the C-terminal of SIN I is able to contact a protein that does specifically bind a short DNA sequence (XBS) in the HO promoter [8]. In this paper we have further pursued the role of this DNA sequence in regulating the transcription of HO, and have shown that the HMGI domain of SINI itself can preferentially bind this sequence, These experiments have demonstrated that the XBS is an important component of the c;sregulatory sequences that participate in the modulation of HO transcription, and that this modulation is accomplished by interactions of SIN i and the SWI/SNF complex at this locus.…”

Section: C~hslommentioning

confidence: 99%

Characterization of a short unique sequence in the yeast HO gene promoter that regulates HO transcription in a SIN1 dependent manner

et al. 1996

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Recently it has become clear that general chromatin proteins as well as sequenco-specifle DNA b!nding protein, s are important in the control of gene expression. SIN1 in Saccharomyces cerevisiae is a chromatln component that regulates the transcription of a family of genes. Previously, we identified a 32 bp unique sequence (here termed XB$) in the promoter of one of those genes, HO, which s~eeifleally binds a protein that interacts with SINI. We also found that this sequence can function as a weak UAS in a beterologous promoter that is dependent on the presence of SINI. Here we report a relationship between the level of llO expreMIon and the presence of the short sequence in situ in the HO gene. By comparing the expression of llO from wild type or XB$ deleted HO promoters, we concluded that XBS serves as a weak UA$ in situ in the HO gene, that it influences HO transcription via the SWIlSNF complex, and that sequences other than the XBS mediate the affect of SINI on HO transcription. In addition, we show that a portion of the SINI protein that has sequence similarity to mammalian HMGI preferentially binds the XBS.

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“…Several hypothetical functional domains in Sin1p have been defined based on genetic, structural, and biochemical considerations (6,9,21,(23)(24)(25). These domains include two HMG1-like domains, (amino acids 26-88 and 98-159), an acidic domain (amino acids 224-304), and a basic C-terminal domain (amino acids 303-333).…”

Section: Sin1p Binds Downstream Of Many Genes Just Upstream Of a Majormentioning

confidence: 99%

Recruitment of mRNA cleavage/polyadenylation machinery by the yeast chromatin protein Sin1p/Spt2p

Hershkovits

Bangio²,

Cohen³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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The yeast chromatin protein Sin1p͞Spt2p has long been studied, but the understanding of its function has remained elusive. The protein has sequence similarity to HMG1, specifically binds crossing DNA structures, and serves as a negative transcriptional regulator of a small family of genes that are activated by the SWI͞SNF chromatin-remodeling complex. Recently, it has been implicated in maintaining the integrity of chromatin during transcription elongation. Here we present experiments whose results indicate that Sin1p͞Spt2 is required for, and is directly involved in, the efficient recruitment of the mRNA cleavage͞polyadenylation complex. This conclusion is based on the following findings: Sin1p͞Spt2 frequently binds specifically downstream of many ORFs but almost always upstream of the first polyadenylation site. It directly interacts with Fir1p, a component of the cleavage͞polyadenylation complex. Disruption of Sin1p͞Spt2p results in foreshortened poly(A) tracts on mRNA. It is synthetically lethal with Cdc73p, which is involved in the recruitment of the complex. This report shows that a chromatin component is involved in 3 end processing of RNA. S in1p is a yeast chromatin nonhistone protein that has beenshown to function as a negative transcriptional regulator of several genes, including Suc2 (1), Ino1 (2), and SSA3 (3). Activity of the HO promoter, as measured from an HO promoter driving a lacZ gene (4), is also affected by Sin1. Sin1 (also known as spt2 in this context) mutants were identified as suppressors of Ty and insertions in the 5Ј noncoding region of the HIS4 gene (5). Because the negative regulation of these genes is overcome by the SW1͞SNF chromatin-remodeling complex (4, 6, 7) and the C-terminal domain of Sin1p interacts with Swi1p (8), it was suggested that a function of SIN1p is to somehow maintain chromatin compaction at specific loci in the chromatin. Peterson et al. (2) found a functional relationship between the C-terminal domain (CTD) of RNA polymerase II and Sin1p, but these data were not pursued further. Sequence analysis of Sin1p showed sequence similarity in two domains to HMG1 (6, 7), a known chromatin protein. Work from our laboratory showed that Sin1p can bind four-way junction and crossing DNA structures (9), supporting the idea that Sin1p binds DNA as it enters and exits the nucleosome.In the context of a global mapping project, Tong et al. (10) reported that there is a synthetic lethal interaction between sin1 and cdc73, a member of the PAF complex. The PAF complex, which accompanies RNA polymerase II during elongation, was shown to have an important function in 3Ј end formation and in polyadenylation (11-13). In addition, a functional interaction was demonstrated between Sin1p and Hpr1p, which is associated with the PAF complex (14).Most recently, evidence has been presented indicating that Sin1p͞Spt2p plays roles in transcription elongation, chromatin structure and genome stability (15). In that study, synthetic lethal interactions were reported between sin1 and paf1, and betwe...

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The C-terminal domain of SIN1 in yeast interacts with a protein that binds the URS1 region of the yeast HO gene

Cited by 8 publications

References 20 publications

Functional Domains of the Yeast Chromatin Protein Sin1p/Spt2p Can Bind Four-way Junction and Crossing DNA Structures

Functional Domains of the Yeast Chromatin Protein Sin1p/Spt2p Can Bind Four-way Junction and Crossing DNA Structures

Characterization of a short unique sequence in the yeast HO gene promoter that regulates HO transcription in a SIN1 dependent manner

Recruitment of mRNA cleavage/polyadenylation machinery by the yeast chromatin protein Sin1p/Spt2p

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