Combination of genomic approaches with functional genetic experiments reveals two modes of repression of yeast middle-phase meiosis genes

BackgroundRegulation of meiosis and sporulation in Saccharomyces cerevisiae is a model for a highly regulated developmental process. Meiosis middle phase transcriptional regulation is governed by two transcription factors: the activator Ndt80 and the repressor Sum1. It has been suggested that the competition between Ndt80 and Sum1 determines the temporal expression of their targets during middle meiosis.ResultsUsing a combination of ChIP-on-chip and expression profiling, we characterized a middle phase transcriptional network and studied the relationship between Ndt80 and Sum1 during middle and late meiosis. While finding a group of genes regulated by both factors in a feed forward loop regulatory motif, our data also revealed a large group of genes regulated solely by Ndt80. Measuring the expression of all Ndt80 target genes in various genetic backgrounds (WT, sum1Δ and MK-ER-Ndt80 strains), allowed us to dissect the exact transcriptional network regulating each gene, which was frequently different than the one inferred from the binding data alone.ConclusionThese results highlight the need to perform detailed genetic experiments to determine the relative contribution of interactions in transcriptional regulatory networks.

[1]  Philipp J. Keller,et al.  Nud1p, the yeast homolog of Centriolin, regulates spindle pole body inheritance in meiosis , 2006, The EMBO journal.

[2]  Giancarlo Castellano,et al.  The involvement of the transcription factor Yin Yang 1 in cancer development and progression , 2009, Cell cycle.

[3]  Aviv Regev,et al.  Dissecting Timing Variability in Yeast Meiosis , 2007, Cell.

[4]  Clifford A. Meyer,et al.  Model-based analysis of tiling-arrays for ChIP-chip , 2006, Proceedings of the National Academy of Sciences.

[5]  E. Fraenkel,et al.  WebMOTIFS: automated discovery, filtering and scoring of DNA sequence motifs using multiple programs and Bayesian approaches , 2007, Environmental health perspectives.

[6]  L. David,et al.  Four Linked Genes Participate in Controlling Sporulation Efficiency in Budding Yeast , 2006, PLoS genetics.

[7]  Larry Kedes,et al.  HES and HERP families: Multiple effectors of the notch signaling pathway , 2003, Journal of cellular physiology.

[8]  Amir Sherman,et al.  Multiple and Distinct Activation and Repression Sequences Mediate the Regulated Transcription of IME1, a Transcriptional Activator of Meiosis-Specific Genes inSaccharomyces cerevisiae , 1998, Molecular and Cellular Biology.

[9]  S. Shen-Orr,et al.  Network motifs: simple building blocks of complex networks. , 2002, Science.

[10]  E. Pichersky,et al.  Rose scent: genomics approach to discovering novel floral fragrance-related genes. , 2002, The Plant cell.

[11]  M. Kunimatsu,et al.  Mei4p coordinates the onset of meiosis I by regulating cdc25+ in fission yeast , 2007, Proceedings of the National Academy of Sciences.

[12]  S. Shen-Orr,et al.  Networks Network Motifs : Simple Building Blocks of Complex , 2002 .

[13]  Sheetal Raithatha,et al.  Phosphorylation and Maximal Activity of Saccharomyces cerevisiae Meiosis-Specific Transcription Factor Ndt80 Is Dependent on Ime2 , 2002, Molecular and Cellular Biology.

[14]  M Wilm,et al.  The S. cerevisiae SET3 complex includes two histone deacetylases, Hos2 and Hst1, and is a meiotic-specific repressor of the sporulation gene program. , 2001, Genes & development.

[15]  Masayuki Yamamoto,et al.  Molecular mechanisms underlying the mitosis–meiosis decision , 2007, Chromosome Research.

[16]  D. Botstein,et al.  The transcriptional program of sporulation in budding yeast. , 1998, Science.

[17]  A. Mitchell,et al.  Bipartite structure of an early meiotic upstream activation sequence from Saccharomyces cerevisiae , 1993, Molecular and cellular biology.

[18]  S. Shen-Orr,et al.  Network motifs in the transcriptional regulation network of Escherichia coli , 2002, Nature Genetics.

[19]  S. Mangan,et al.  Structure and function of the feed-forward loop network motif , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Naama Barkai,et al.  Modulation of the transcription regulatory program in yeast cells committed to sporulation , 2006, Genome Biology.

[21]  H. E. Smith,et al.  Role of IME1 expression in regulation of meiosis in Saccharomyces cerevisiae , 1990, Molecular and cellular biology.

[22]  John J. Wyrick,et al.  Genome-wide location and function of DNA binding proteins. , 2000, Science.

[23]  Olga G. Troyanskaya,et al.  GOLEM: an interactive graph-based gene-ontology navigation and analysis tool , 2006, BMC Bioinformatics.

[24]  Y. Kassir,et al.  Genetic regulation of differentiation towards meiosis in the yeast Saccharomyces cerevisiae. , 1989, Genome.

[25]  G. Simchen Commitment to meiosis: what determines the mode of division in budding yeast? , 2009, BioEssays : news and reviews in molecular, cellular and developmental biology.

[26]  Chao Zhang,et al.  Control of landmark events in meiosis by the CDK Cdc28 and the meiosis-specific kinase Ime2. , 2003, Genes & development.

[27]  Nicola J. Rinaldi,et al.  Serial Regulation of Transcriptional Regulators in the Yeast Cell Cycle , 2001, Cell.

[28]  R. D. Gietz,et al.  Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. , 2002, Methods in enzymology.

[29]  A. Mitchell Control of meiotic gene expression in Saccharomyces cerevisiae. , 1994, Microbiological reviews.

[30]  J. Pak,et al.  Regulation of the Premiddle and Middle Phases of Expression of the NDT80 Gene during Sporulation of Saccharomyces cerevisiae , 2002, Molecular and Cellular Biology.

[31]  G. Simchen,et al.  Sporulation in Saccharomyces cerevisiae: premeiotic DNA synthesis, readiness and commitment. , 1972, Experimental cell research.

[32]  S. Chu,et al.  Gametogenesis in yeast is regulated by a transcriptional cascade dependent on Ndt80. , 1998, Molecular cell.

[33]  A. Vershon,et al.  Sum1 and Hst1 repress middle sporulation‐specific gene expression during mitosis in Saccharomyces cerevisiae , 1999, The EMBO journal.

[34]  Amir Sherman,et al.  Multiple and Distinct Activation and Repression Sequences Mediate the Regulated Transcription of IME1, a Transcriptional Activator of Meiosis-Specific Genes in Saccharomyces cerevisiae , 1998, Molecular and Cellular Biology.

[35]  Nicola J. Rinaldi,et al.  Transcriptional regulatory code of a eukaryotic genome , 2004, Nature.

[36]  A. Amon,et al.  Meiosis I Is Established through Division-Specific Translational Control of a Cyclin , 2008, Cell.

[37]  Sanjit K. Mitra,et al.  Optimized LOWESS normalization parameter selection for DNA microarray data , 2004, BMC Bioinformatics.

[38]  Galit Shenhar,et al.  Transcriptional regulation of meiosis in budding yeast. , 2003, International review of cytology.

[39]  M. J. Mallory,et al.  Meiosis-specific destruction of the Ume6p repressor by the Cdc20-directed APC/C. , 2007, Molecular cell.

[40]  Philipp J. Keller,et al.  Spore number control and breeding in Saccharomyces cerevisiae , 2005, The Journal of cell biology.

[41]  K. Benjamin,et al.  Sum1 and Ndt80 Proteins Compete for Binding to Middle Sporulation Element Sequences That Control Meiotic Gene Expression , 2003, Molecular and Cellular Biology.

[42]  Ronald W. Davis,et al.  The core meiotic transcriptome in budding yeasts , 2000, Nature Genetics.

[43]  Emmitt R. Jolly,et al.  Inference of combinatorial regulation in yeast transcriptional networks: a case study of sporulation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[44]  P. T. Magee,et al.  Isolation of genes expressed preferentially during sporulation in the yeast Saccharomyces cerevisiae. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[45]  L. Xu,et al.  NDT80, a meiosis-specific gene required for exit from pachytene in Saccharomyces cerevisiae , 1995, Molecular and cellular biology.

[46]  U. Alon Network motifs: theory and experimental approaches , 2007, Nature Reviews Genetics.

[47]  A. Vershon,et al.  The pachytene checkpoint in Saccharomyces cerevisiae requires the Sum1 transcriptional repressor , 2000, The EMBO journal.

[48]  Motonao Nakamura,et al.  Positive and negative elements upstream of the meiosis-specific glucoamylase gene in Saccbaromyces cerevisiae , 1991, Molecular and General Genetics MGG.

[49]  Y. Shiio,et al.  Epitope tagging. , 1995, Methods in enzymology.

[50]  K. Siegers,et al.  Epitope tagging of yeast genes using a PCR‐based strategy: more tags and improved practical routines , 1999, Yeast.

[51]  R. Akada,et al.  Positive and negative elements upstream of the meiosis-specific glucoamylase gene in Saccharomyces cerevisiae. , 1991, Molecular & general genetics : MGG.

[52]  R. Rothstein Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. , 1991, Methods in enzymology.

[53]  I. Simon,et al.  Genome-wide transcriptional analysis of the human cell cycle identifies genes differentially regulated in normal and cancer cells , 2008, Proceedings of the National Academy of Sciences.

[54]  I. Simon,et al.  Deconvolving cell cycle expression data with complementary information , 2004, ISMB/ECCB.

[55]  Docent Jarmo Jääskeläinen,et al.  Genetic Regulation , 2008 .