title: Characterizing the roles of Met31 and Met32 in coordinating Met4-activated transcription in the absence of Met30.
abstract: Yeast sulfur metabolism is transcriptionally regulated by the activator Met4. Met4 lacks DNA-binding ability and relies on interactions with Met31 and Met32, paralogous proteins that bind the same cis-regulatory element, to activate its targets. Although Met31 and Met32 are redundant for growth in the absence of methionine, studies indicate that Met32 has a prominent role over Met31 when Met30, a negative regulator of Met4 and Met32, is inactive. To characterize different roles of Met31 and Met32 in coordinating Met4-activated transcription, we examined transcription in strains lacking either Met31 or Met32 upon Met4 induction in the absence of Met30. Microarray analysis revealed that transcripts involved in sulfate assimilation and sulfonate metabolism were dramatically decreased in met32Δ cells compared to its wild-type and met31Δ counterparts. Despite this difference, both met31Δ and met32Δ cells used inorganic sulfur compounds and sulfonates as sole sulfur sources in minimal media when Met30 was present. This discrepancy may be explained by differential binding of Met31 to Cbf1-dependent promoters between these two conditions. In the absence of Met30, genome-wide chromatin immunoprecipitation analyses found that Met32 bound all Met4-bound targets, supporting Met32 as the main platform for Met4 recruitment. Finally, Met31 and Met32 levels were differentially regulated, with Met32 levels mimicking the profile for active Met4. These different properties of Met32 likely contribute to its prominent role in Met4-activated transcription when Met30 is absent.
authors.affiliation: Department of Biological Sciences, University of Wisconsin-Parkside, Kenosha, WI 53144, USA.
authors.forename: Emilio
authors.initials: E
authors.lastname: Carrillo
authors.forename: Giora
authors.initials: G
authors.lastname: Ben-Ari
authors.forename: Jan
authors.initials: J
authors.lastname: Wildenhain
authors.forename: Mike
authors.initials: M
authors.lastname: Tyers
authors.forename: Dilon
authors.initials: D
authors.lastname: Grammentz
authors.forename: Traci A
authors.initials: TA
authors.lastname: Lee
mesh_terms: D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000378:metabolism*; D001665:Binding Sites; D016000:Cluster Analysis; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D044783:F-Box Proteins / Q000235:genetics; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D013451:Sulfonic Acids / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D043743:Ubiquitin-Protein Ligase Complexes / Q000172:deficiency* / Q000235:genetics
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D044783:F-Box Proteins; C470285:MET30 protein, S cerevisiae; C073675:MET4 protein, S cerevisiae; C106764:Met31 protein, S cerevisiae; C106765:Met32 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013451:Sulfonic Acids; D014157:Transcription Factors; D043743:Ubiquitin-Protein Ligase Complexes