Structure of the sporulation-specific transcription factor Ndt80 bound to DNA

Lamoureux, Jason S.; Stuart, David A.; Tsang, Roger Y.; Wu, Cynthia; Glover, J. N. Mark

doi:10.1093/emboj/cdf572

Cited by 63 publications

(88 citation statements)

References 53 publications

(91 reference statements)

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“…Although this element is functional in vitro (Lamoureux et al 2002), the regulation of DIT1 is not fully understood and is a good illustration of how difficult it is to localize regulation to specific binding sites (see supplemental methods at http:/ /www.genetics.org/supplemental/). For our library, we took 27 bp surrounding this MSE site and called it MSE-DIT.…”

Section: Resultsmentioning

confidence: 99%

Gene Expression From Random Libraries of Yeast Promoters

et al. 2006

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Genomewide techniques to assay gene expression and transcription factor binding are in widespread use, but are far from providing predictive rules for the function of regulatory DNA. To investigate more intensively the grammar rules for active regulatory sequence, we made libraries from random ligations of a very restricted set of sequences. Working with the yeast Saccharomyces cerevisiae, we developed a novel screen based on the sensitivity of ascospores lacking dityrosine to treatment with lytic enzymes. We tested two separate libraries built by random ligation of a single type of activator site either for a wellcharacterized sporulation factor, Ndt80, or for a new sporulation-specific regulatory site that we identified and several neutral spacer elements. This selective system achieved up to 1:10 4 enrichment of the artificial sequences that were active during sporulation, allowing a high-throughput analysis of large libraries of synthetic promoters. This is not practical with methods involving direct screening for expression, such as those based on fluorescent reporters. There were very few false positives, since active promoters always passed the screen when retested. The survival rate of our libraries containing roughly equal numbers of spacers and activators was a few percent that of libraries made from activators alone. The sequences of 100 examples of active and inactive promoters could not be distinguished by simple binary rules; instead, the best model for the data was a linear regression fit of a quantitative measure of gene activity to multiple features of the regulatory sequence.

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Section: Resultsmentioning

confidence: 99%

Gene Expression From Random Libraries of Yeast Promoters

et al. 2006

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“…We refer to this element as MSE2. Although this element does contain a mismatch to the consensus MSE at position 3 (C to T), mutational studies suggest that this alteration may weaken but is unlikely to ablate the ability of Ndt80 to bind to this sequence in vitro or in vivo (Lamoureux et al, 2002;Pierce et al, 2003).…”

Section: Inactivation Of the Putative Mse1 Does Not Alter Clb5 Expresmentioning

confidence: 99%

“…Ndt80 is an Ig-fold transcription factor that activates the expression of >150 middle sporulation genes by binding to a regulatory DNA sequence known as the middle sporulation element (MSE) found in the promoters of many of these genes Lamoureux et al, 2002;Primig et al, 2000). The NDT80 gene displays a bimodal pattern of transcription.…”

Section: Introductionmentioning

confidence: 99%

The Saccharomyces cerevisiae CLB5 promoter contains two middle sporulation elements (MSEs) that are differentially regulated during sporulation

Raithatha

Stuart

2008

Yeast

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The B-type cyclins Clb5 and Clb6 are essential activators of DNA replication during sporulation in Saccharomyces cerevisiae. The expression of CLB5 is maximally induced during the middle phase of sporulation by the transcription factor Ndt80. We have performed an analysis of the CLB5 promoter and have identified two middle sporulation elements (MSEs) that act as binding sites for Ndt80. Although both MSE sequences bind Ndt80 in vitro, they display differential effectiveness in their ability to function as cis-acting regulatory sequences in vivo. Mutation of both MSE sequences in the CLB5 promoter profoundly reduces the induction of CLB5 transcription during the middle phase of sporulation but results in no obvious defect in progression through meiosis and sporulation, implying that the Ndt80-dependent induction of CLB5 is not required for effective DNA replication or chromosome division.

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“…Ndt80 belongs to the Ig-fold family of transcription factors (Lamoureux et al 2002;Montano et al 2002) and regulates the prophase-tometaphase transition in S. cerevisiae (Chu and Herskowitz 1998;Hepworth et al 1998;Tung et al 2000;Pak and Segall 2002b). Ndt80 controls the expression of $150-200 genes that regulate meiotic progression and spore formation (Chu and Herskowitz 1998;).…”

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confidence: 99%

Meiotic Regulators Ndt80 and Ime2 Have Different Roles in Saccharomyces and Neurospora

Hutchison

Glass

2010

Genetics

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Meiosis is a highly regulated process in eukaryotic species. The filamentous fungus Neurospora crassa has been shown to be missing homologs of a number of meiotic initiation genes conserved in Saccharomyces cerevisiae, but has three homologs of the well-characterized middle meiotic transcriptional regulator NDT80. In this study, we evaluated the role of all three NDT80 homologs in the formation of female reproductive structures, sexual development, and meiosis. We found that none of the NDT80 homologs were required for meiosis and that even the triple mutant was unaffected. However, strains containing mutations in NCU09915 (fsd-1) were defective in female sexual development and ascospore maturation. vib-1 was a major regulator of protoperithecial development in N. crassa, and double mutants carrying deletions of both vib-1 (NCU03725) and fsd-1 exhibited a synergistic effect on the timing of female reproductive structure (protoperithecia) formation. We further evaluated the role of the N. crassa homolog of IME2, a kinase involved in initiation of meiosis in S. cerevisiae. Strains containing mutations in ime-2 showed unregulated development of protoperithecia. Genetic analysis indicated that mutations in vib-1 were epistatic to ime-2, suggesting that IME-2 may negatively regulate VIB-1 activity. Our data indicate that the IME2/NDT80 pathway is not involved in meiosis in N. crassa, but rather regulates the formation of female reproductive structures.

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Structure of the sporulation-specific transcription factor Ndt80 bound to DNA

Cited by 63 publications

References 53 publications

Gene Expression From Random Libraries of Yeast Promoters

Gene Expression From Random Libraries of Yeast Promoters

The Saccharomyces cerevisiae CLB5 promoter contains two middle sporulation elements (MSEs) that are differentially regulated during sporulation

Meiotic Regulators Ndt80 and Ime2 Have Different Roles in Saccharomyces and Neurospora

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