Green Fluorescent Protein-Dal80p Illuminates up to 16 Distinct Foci That Colocalize with and Exhibit the Same Behavior as Chromosomal DNA Proceeding through the Cell Cycle of
            <i>Saccharomyces cerevisiae</i>

Distler, Mackenzie; Kulkarni, Ajit A.; Rai, Rajendra; Cooper, Terrance G.

doi:10.1128/jb.183.15.4636-4642.2001

Cited by 5 publications

(3 citation statements)

References 28 publications

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“…In order to determine the genome-wide occupancy of a GATA factor in yeast, our rationale was to choose Dal80 as it is known to be highly expressed in derepressing conditions and forms chromosome foci when tagged by GFP [ 56 ]. We grew yeast cells in proline-containing medium and performed a ChIP-Seq analysis using a Dal80-Myc 13 -tagged strain and the isogenic untagged strain, as a control ( Fig 1A ), after ensuring that the Myc 13 -tagged form of Dal80 was functional ( S1A Fig ).…”

Section: Resultsmentioning

confidence: 99%

Transcription-dependent spreading of the Dal80 yeast GATA factor across the body of highly expressed genes

et al. 2019

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GATA transcription factors are highly conserved among eukaryotes and play roles in transcription of genes implicated in cancer progression and hematopoiesis. However, although their consensus binding sites have been well defined in vitro, the in vivo selectivity for recognition by GATA factors remains poorly characterized. Using ChIP-Seq, we identified the Dal80 GATA factor targets in yeast. Our data reveal Dal80 binding to a large set of promoters, sometimes independently of GATA sites, correlating with nitrogen- and/or Dal80-sensitive gene expression. Strikingly, Dal80 was also detected across the body of promoter-bound genes, correlating with high expression. Mechanistic single-gene experiments showed that Dal80 spreading across gene bodies requires active transcription. Consistently, Dal80 co-immunoprecipitated with the initiating and post-initiation forms of RNA Polymerase II. Our work suggests that GATA factors could play dual, synergistic roles during transcription initiation and post-initiation steps, promoting efficient remodeling of the gene expression program in response to environmental changes.

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Section: Resultsmentioning

confidence: 99%

Transcription-dependent spreading of the Dal80 yeast GATA factor across the body of highly expressed genes

et al. 2019

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“…Cells transformed with a GFP-DAL80 plasmid exhibit fluorescent foci that co-localize with 4,6-diamidino-2-phenylindole-positive material and follow DNA movement through the cell cycle (45). High resolution confocal and deconvolution microscopy delineate up to 16 distinct foci, which correlates with the number of S. cerevisiae chromosomes (45).…”

Section: Gfp-gln3p Occasionally Exhibits Intranuclear Localization Simentioning

confidence: 99%

“…High resolution confocal and deconvolution microscopy delineate up to 16 distinct foci, which correlates with the number of S. cerevisiae chromosomes (45). DAL80 encodes a repressor protein, which binds to some of the same GATA sequences as Gln3p (10,32,(46)(47)(48).…”

Section: Gfp-gln3p Occasionally Exhibits Intranuclear Localization Simentioning

confidence: 99%

Gln3p Nuclear Localization and Interaction with Ure2p inSaccharomyces cerevisiae

Kulkarni

Abulhamd

Rai

et al. 2001

Journal of Biological Chemistry

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Gln3p is one of two well characterized GATA family transcriptional activation factors whose function is regulated by the nitrogen supply of the cell. When nitrogen is limiting, Gln3p and Gat1p are concentrated in the nucleus where they bind GATA sequences upstream of nitrogen catabolite repression (NCR)-sensitive genes and activate their transcription. Conversely, in excess nitrogen, these GATA sequences are unoccupied by Gln3p and Gat1p because these transcription activators are excluded from the nucleus. Ure2p binds to Gln3p and Gat1p and is required for NCR-sensitive transcription to be repressed and for nuclear exclusion of these transcription factors. Here we show the following. (i) Gln3p residues 344-365 are required for nuclear localization. (ii) Replacing Ser-344, Ser-347, and Ser-355 with alanines has minimal effects on GFP-Gln3p localization. However, replacing Gln3p Ser-344, Ser-347, and Ser-355 with aspartates results in significant loss of its ability to be concentrated in the nucleus. (iii) N and C termini of the Gln3p region required for it to complex with Ure2p and be excluded from the nucleus are between residues 1-103 and 301-365, respectively. (iv) N and C termini of the Ure2p region required for it to interact with Gln3p are situated between residues 101-151 and 330-346, respectively. (v) Loss of Ure2p residues participating in either dimer or prion formation diminishes its ability to carry out NCR-sensitive regulation of Gln3p activity.Saccharomyces cerevisiae is increasingly used as a model to identify the functions of mammalian proteins as well as how their production and activities are regulated and integrated. One of the gene families shared by S. cerevisiae and higher eukaryotes is the GATA family of DNA-binding proteins. In animal cells, GATA family proteins were originally shown to be responsible for regulating globin gene expression (1). However, they are now known to regulate a diverse set of developmental functions (2, 3). In yeast, the GATA family proteins Gln3p, Gat1p/Nil1p, Dal80p, Deh1p/Gzf3p have been studied as the main regulators of nitrogen catabolic gene expression (4-6), and recently their regulatory functions also been shown to be far more diverse (7,8). Gln3p and Gat1p are transcriptional activators, whereas Dal80p and Deh1p repress transcription by competing with these activators for binding to their target GATA sequences (4)(5)(6)(9)(10)(11) HHS Public Access Author Manuscript Author ManuscriptAuthor Manuscript Author ManuscriptGln3p and Gat1p function is also dramatically regulated by the nitrogen supply of the cell. When nitrogen is limiting, Gln3p and Gat1p are concentrated in the nucleus, bind to the GATA sequences of their target promoters, and activate transcription (8,9,(12)(13)(14)(15). On the other hand, when nitrogen is in excess, Gln3p and Gat1p are excluded from the nucleus, and nitrogen catabolite repression (NCR) 1 -sensitive gene expression is repressed. Nuclear exclusion of Gln3p and Gat1p requires Ure2p, the first NCR regulator identified (16...

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Current Awareness

2001

Yeast

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In order to keep subscribers up‐to‐date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly‐published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals ‐ search completed 5th. Sep. 2001)

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Green Fluorescent Protein-Dal80p Illuminates up to 16 Distinct Foci That Colocalize with and Exhibit the Same Behavior as Chromosomal DNA Proceeding through the Cell Cycle of Saccharomyces cerevisiae

Cited by 5 publications

References 28 publications

Transcription-dependent spreading of the Dal80 yeast GATA factor across the body of highly expressed genes

Transcription-dependent spreading of the Dal80 yeast GATA factor across the body of highly expressed genes

Gln3p Nuclear Localization and Interaction with Ure2p inSaccharomyces cerevisiae

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