Similar upstream regulatory elements of genes that encode the two largest subunits of RNA polymerase II in Saccharomyces cerevisiae.

We have determined the location of cis-acting elements that are important for the expression of RPO21 and RPO22, genes that encode the two largest subunits of RNA polymerase II (RNAPII) in Saccharomyces cerevisiae. A series of 5'-end deletions and nucleotide substitutions in the upstream regions of RPO21 and RPO22 were tested for their effect on the expression of lacZ fusions of these genes. Deletion of sequences from -723 to -693 in RPO21, which disrupted two Reb1p-binding sites and an Abf1p-binding site, resulted in a 10-fold decrease in expression. A T-rich region downstream of these sites was also important for expression. Deletion of sequences from -437 to -392 in the RPO22-upstream, which resulted in a 30-fold decrease in expression, indicated that the Reb1p- and Abf1p-binding sites in this region were important for RPO22 expression, as was a T-rich sequence immediately downstream of these sites. The RPO21 and RPO22 upstream regions were capable of interacting in vitro (gel-mobility-shift assays) with Reb1p and Abf1p. The similarities in the type and organization of elements in the upstream regions of RPO21 and RPO22 suggest that expression of these genes may be regulated coordinately.

[1]  V. Iyer,et al.  Poly(dA:dT), a ubiquitous promoter element that stimulates transcription via its intrinsic DNA structure. , 1995, The EMBO journal.

[2]  J. Broach,et al.  Nutrient availability and the RAS/cyclic AMP pathway both induce expression of ribosomal protein genes in Saccharomyces cerevisiae but by different mechanisms , 1995, Molecular and cellular biology.

[3]  W. H. Mager,et al.  Transcription activation of yeast ribosomal protein genes requires additional elements apart from binding sites for Abf1p or Rap1p. , 1995, Nucleic acids research.

[4]  D. Shore,et al.  RAP1: a protean regulator in yeast. , 1994, Trends in genetics : TIG.

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

[6]  C. Brandl,et al.  Defining the sequence specificity of the Saccharomyces cerevisiae DNA binding protein REB1p by selecting binding sites from random‐sequence oligonucleotides , 1994, Yeast.

[7]  M. Holland,et al.  The upstream repression sequence from the yeast enolase gene ENO1 is a complex regulatory element that binds multiple trans-acting factors including REB1. , 1994, The Journal of biological chemistry.

[8]  K. Struhl,et al.  Protein kinase A mediates growth-regulated expression of yeast ribosomal protein genes by modulating RAP1 transcriptional activity , 1994, Molecular and cellular biology.

[9]  H. Stunnenberg,et al.  Improved method for PCR-mediated site-directed mutagenesis. , 1994, Nucleic acids research.

[10]  J. Workman,et al.  Multiple functions of nucleosomes and regulatory factors in transcription. , 1993, Trends in biochemical sciences.

[11]  Q. Ju,et al.  A bipartite DNA-binding domain in yeast Reb1p , 1993, Molecular and cellular biology.

[12]  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.

[13]  S. Elsasser,et al.  Role of multifunctional autonomously replicating sequence binding factor 1 in the initiation of DNA replication and transcriptional control in Saccharomyces cerevisiae , 1992, Molecular and cellular biology.

[14]  D. E. Griffiths,et al.  DMSO-enhanced whole cell yeast transformation. , 1991, Nucleic acids research.

[15]  R. Young,et al.  RNA polymerase II. , 1991, Annual review of biochemistry.

[16]  A. Chambers,et al.  ARS binding factor 1 binds adjacent to RAP1 at the UASs of the yeast glycolytic genes PGK and PYK1. , 1990, Nucleic acids research.

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

[18]  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.

[19]  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.

[20]  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.

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

[22]  D. Chasman,et al.  A yeast protein that influences the chromatin structure of UASG and functions as a powerful auxiliary gene activator. , 1990, Genes & development.

[23]  H. Wang,et al.  Identification of a Saccharomyces cerevisiae DNA-binding protein involved in transcriptional regulation , 1990, Molecular and cellular biology.

[24]  R. Kornberg,et al.  A yeast ARS-binding protein activates transcription synergistically in combination with other weak activating factors , 1990, Molecular and cellular biology.

[25]  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.

[26]  B. Morrow,et al.  Proteins that bind to the yeast rDNA enhancer. , 1989, The Journal of biological chemistry.

[27]  K. Struhl.  Molecular mechanisms of transcriptional regulation in yeast. , 1990, Annual review of biochemistry.

[28]  R. Kornberg,et al.  Statistical positioning of nucleosomes by specific protein-binding to an upstream activating sequence in yeast. , 1988, Journal of molecular biology.

[29]  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.

[30]  J. Rine,et al.  Roles of two DNA‐binding factors in replication, segregation and transcriptional repression mediated by a yeast silencer. , 1988, The EMBO journal.

[31]  R. Pearlman,et al.  Transposable element-mediated enhancement of gene expression in Saccharomyces cerevisiae involves sequence-specific binding of a trans-acting factor , 1988, Molecular and cellular biology.

[32]  R. Planta,et al.  Control of ribosome biogenesis in yeast. , 1988, Trends in genetics : TIG.

[33]  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.

[34]  R. Young,et al.  Eucaryotic RNA polymerase conditional mutant that rapidly ceases mRNA synthesis , 1987, Molecular and cellular biology.

[35]  R. Young,et al.  Prokaryotic and eukaryotic RNA polymerases have homologous core subunits. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[36]  Michael Shales,et al.  Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases , 1985, Cell.

[37]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[38]  D. Crothers,et al.  Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. , 1981, Nucleic acids research.

[39]  L. Guarente,et al.  Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[40]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[41]  A. Ishihama,et al.  Genetics of bacterial RNA polymerases. , 1979, Annual review of genetics.

[42]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[43]  B. Hall,et al.  Transcription in yeast: separation and properties of multiple FNA polymerases. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .