Participation of the yeast activator Abf1 in meiosis-specific expression of the HOP1 gene

The meiosis-specific gene HOP1, which encodes a component of the synaptonemal complex, is controlled through two regulatory elements, UASH and URS1H. Sites similar to URS1H have been identified in the promoter region of virtually every early meiosis-specific gene, as well as in many promoters of nonmeiotic genes, and it has been shown that the proteins that bind to this site function to regulate meiotic and nonmeiotic transcription. Sites similar to the UASH site have been found in a number of meiotic and nonmeiotic genes as well. Since it has been shown that UASH functions as an activator site in vegetative haploid cells, it seemed likely that the factors binding to this site regulate both meiotic and nonmeiotic transcription. We purified the factor binding to the UASH element of the HOP1 promoter. Sequence analysis identified the protein as Abf1 (autonomously replicating sequence-binding factor 1), a multifunctional protein involved in DNA replication, silencing, and transcriptional regulation. We show by mutational analysis of the UASH site, that positions outside of the proposed UASH consensus sequence (TNTGN[A/T]GT) are required for DNA binding in vitro and transcriptional activation in vivo. A new UASH consensus sequence derived from this mutational analysis closely matches a consensus Abf1 binding site. We also show that an Abf1 site from a nonmeiotic gene can replace the function of the UASH site in the HOP1 promoter. Taken together, these results show that Abf1 functions to regulate meiotic gene expression.

[1]  B. Stillman,et al.  Replication factor-A from Saccharomyces cerevisiae is encoded by three essential genes coordinately expressed at S phase. , 1991, Genes & development.

[2]  L. McBroom,et al.  Contacts of the ABF1 protein of Saccharomyces cerevisiae with a DNA binding site at MATa. , 1994, The Journal of biological chemistry.

[3]  M. Bhattacharyya,et al.  The SPR3 gene encodes a sporulation-specific homologue of the yeast CDC3/10/11/12 family of bud neck microfilaments and is regulated by ABFI. , 1995, Gene.

[4]  J. Rine,et al.  Roles of ABF1, NPL3, and YCL54 in silencing in Saccharomyces cerevisiae. , 1995, Genetics.

[5]  A. Sentenac,et al.  The ABF1 factor is the transcriptional activator of the L2 ribosomal protein genes in Saccharomyces cerevisiae , 1990, Molecular and cellular biology.

[6]  T. Cooper,et al.  Ubiquitous upstream repression sequences control activation of the inducible arginase gene in yeast. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  R. O. Poyton,et al.  ABF1 is a phosphoprotein and plays a role in carbon source control of COX6 transcription in Saccharomyces cerevisiae , 1992, Molecular and cellular biology.

[8]  T. Cooper,et al.  Saccharomyces cerevisiae BUF protein binds to sequences participating in DNA replication in addition to those mediating transcriptional repression (URS1) and activation , 1993, Molecular and cellular biology.

[9]  J. Buhler,et al.  ABF1 binding sites in yeast RNA polymerase genes. , 1990, The Journal of biological chemistry.

[10]  R. E. Esposito,et al.  Meiosis and Ascospore Development , 1981 .

[11]  J. Rine,et al.  Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae , 1988, Molecular and cellular biology.

[12]  N. M. Hollingsworth,et al.  The HOP1 gene encodes a meiosis-specific component of yeast chromosomes , 1990, Cell.

[13]  M. Holland,et al.  Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription , 1990, Molecular and cellular biology.

[14]  K. Hamil,et al.  Constitutive transcription of yeast ribosomal protein gene TCM1 is promoted by uncommon cis- and trans-acting elements , 1988, Molecular and cellular biology.

[15]  B. Stillman,et al.  A yeast chromosomal origin of DNA replication defined by multiple functional elements. , 1992, Science.

[16]  D. Shore,et al.  Identification of silencer binding proteins from yeast: possible roles in SIR control and DNA replication , 1987, The EMBO journal.

[17]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[18]  L. Samson,et al.  Replication protein A binds to regulatory elements in yeast DNA repair and DNA metabolism genes. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[19]  T. Cooper,et al.  Participation of ABF-1 protein in expression of the Saccharomyces cerevisiae CAR1 gene , 1991, Journal of bacteriology.

[20]  H. E. Smith,et al.  A transcriptional cascade governs entry into meiosis in Saccharomyces cerevisiae , 1989, Molecular and cellular biology.

[21]  H. E. Smith,et al.  Positive control of sporulation-specific genes by the IME1 and IME2 products in Saccharomyces cerevisiae , 1990, Molecular and cellular biology.

[22]  A. Mitchell,et al.  Stimulation of later functions of the yeast meiotic protein kinase Ime2p by the IDS2 gene product , 1995, Molecular and cellular biology.

[23]  L. McBroom,et al.  DNA bending by Saccharomyces cerevisiae ABF1 and its proteolytic fragments. , 1994, The Journal of biological chemistry.

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

[25]  R. Padmore,et al.  Temporal comparison of recombination and synaptonemal complex formation during meiosis in S. cerevisiae , 1991, Cell.

[26]  M. Clancy,et al.  An RME1-independent pathway for sporulation control in Saccharomyces cerevisiae acts through IME1 transcript accumulation. , 1990, Genetics.

[27]  A. Mitchell,et al.  The yeast MCK1 gene encodes a protein kinase homolog that activates early meiotic gene expression. , 1991, Genes & development.

[28]  T. Cooper,et al.  The yeast UME6 gene product is required for transcriptional repression mediated by the CAR1 URS1 repressor binding site. , 1992, Nucleic acids research.

[29]  E. Winnacker,et al.  Isolation and DNA‐binding characteristics of a protein involved in transcription activation of two divergently transcribed, essential yeast genes. , 1989, The EMBO journal.

[30]  T. Werner,et al.  MatInd and MatInspector: new fast and versatile tools for detection of consensus matches in nucleotide sequence data. , 1995, Nucleic acids research.

[31]  J. Diffley,et al.  Protein-DNA interactions at a yeast replication origin , 1992, Nature.

[32]  M. Künzler,et al.  Activation and repression of the yeast ARO3 gene by global transcription factors , 1995, Molecular microbiology.

[33]  H. Tabak,et al.  The upstream region of the FOX3 gene encoding peroxisomal 3-oxoacyl-coenzyme A thiolase in Saccharomyces cerevisiae contains ABF1- and replication protein A-binding sites that participate in its regulation by glucose repression , 1995, Molecular and cellular biology.

[34]  W. H. Mager,et al.  An ARS/silencer binding factor also activates two ribosomal protein genes in yeast. , 1989, Nucleic acids research.

[35]  L. Grivell,et al.  Yeast general transcription factor GFI: sequence requirements for binding to DNA and evolutionary conservation. , 1990, Nucleic acids research.

[36]  K. Oegema,et al.  The gene encoding ARS-binding factor I is essential for the viability of yeast. , 1989, Genes & development.

[37]  M. Holland,et al.  Sequences within an upstream activation site in the yeast enolase gene ENO2 modulate repression of ENO2 expression in strains carrying a null mutation in the positive regulatory gene GCR1 , 1990, Molecular and cellular biology.

[38]  C. Szent‐Gyorgyi.  A bipartite operator interacts with a heat shock element to mediate early meiotic induction of Saccharomyces cerevisiae HSP82 , 1995, Molecular and cellular biology.

[39]  J. Broach,et al.  The Molecular biology of the yeast saccharomyces, life cycle and inheritance , 1981 .

[40]  R. E. Esposito,et al.  Identification of negative regulatory genes that govern the expression of early meiotic genes in yeast. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

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

[42]  R. Kolodner,et al.  Characterization of DNA-binding and strand-exchange stimulation properties of y-RPA, a yeast single-strand-DNA-binding protein. , 1992, Journal of molecular biology.

[43]  R. E. Esposito,et al.  UME6 is a key regulator of nitrogen repression and meiotic development. , 1994, Genes & development.

[44]  R. E. Esposito,et al.  Nucleotide sequence and promoter analysis of SPO13, a meiosis-specific gene of Saccharomyces cerevisiae. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[45]  A. Mitchell,et al.  Positive control of yeast meiotic genes by the negative regulator UME6 , 1995, Molecular and cellular biology.

[46]  C. Keleher,et al.  The yeast cell-type-specific repressor α2 acts cooperatively with a non-cell-type-specific protein , 1988, Cell.

[47]  Purification of the heteromeric protein binding to the URS1 transcriptional repression site in Saccharomyces cerevisiae. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[48]  I. Herskowitz,et al.  Activation of meiosis and sporulation by repression of the RME1 product in yeast , 1986, Nature.

[49]  N. M. Hollingsworth,et al.  HOP1: a yeast meiotic pairing gene. , 1989, Genetics.

[50]  K. Sweder,et al.  Purification and characterization of proteins that bind to yeast ARSs. , 1988, The Journal of biological chemistry.

[51]  J. Diffley,et al.  Similarity between the transcriptional silencer binding proteins ABF1 and RAP1. , 1989, Science.