A general upstream binding factor for genes of the yeast translational apparatus.

Glycolytic gene expression in Saccharomyces cerevisiae is thought to be activated by the GCR and TUF proteins. We tested the hypothesis that GCR function is mediated by TUF/GRF/RAP binding sites (UASRPG elements). We found that UASRPG-dependent activation of a heterologous gene and transcription of ADH1, TEF1, TEF2, and RP59 were sensitive to GCR1 disruption. GCR is not required for TUF/GRF/RAP expression or in vitro DNA-binding activity.

mentioning

confidence: 99%

mentioning

confidence: 99%

Efficient transcription of the glycolytic gene ADH1 and three translational component genes requires the GCR1 product, which can act through TUF/GRF/RAP binding sites.

Santangelo¹,

Tornow²

1990

“…For methylation interference, 60,000 cpm of the DNA probes and 3 ,ug of pSP65 DNA were incubated with 2 jig of proteins from fraction 80 and treated as described by Carnevali et al (5); for DNase I footprinting, 10,000 cpm of the DNA probes and 3 ,ug of pSP65 DNA were incubated with 3 jig of proteins from fraction 80 and treated as described by Huet et al. (14). On the coding strand of the L2A promoter, a guanine at position -179 was a major contact point of the factor; a second guanine at position -170 was a minor contact point (Fig.…”

mentioning

confidence: 99%

The ABF1 factor is the transcriptional activator of the L2 ribosomal protein genes in Saccharomyces cerevisiae.

Seta¹,

Ciafrè²,

Marck³

et al. 1990

100

The same factor, ABF1, binds to the promoters of the two gene copies (L2A and L2B) coding for the ribosomal protein L2 in Saccharomyces cerevisiae. In vitro binding experiments and in vivo functional analysis showed that the different affinities of the L2A and L2B promoters for the ABF1 factor are responsible for the differential transcriptional activities of the two gene copies. The presence of ABF1-binding sites in front of many housekeeping genes suggests a general role for ABF1 in the regulation of gene activity.

“…* We have recently identified by detailed mutational analysis three sequence motifs upstream of the RP39A gene necessary for its transcription (58). These sequences are present in the same 5'-to-3' orientation upstream of 18 of the 20 cloned ribosomal protein genes (38,58,66), as well as the translational elongation factor genes TEFI and TEF2 (23).…”

mentioning

confidence: 99%

Structure and expression of the Saccharomyces cerevisiae CRY1 gene: a highly conserved ribosomal protein gene.

Larkin¹,

Thompson²,

Woolford³

1987

The Saccharomyces cerevisiae CRY] gene encodes ribosomal protein rp59, a component of the 40S ribosomal subunit. Mutations in CRY] can confer resistance to the alkaloid cryptopleurine, an inhibitor of the elongation step of translation. The nucleotide sequence of the cloned CRY] gene was determined. The predicted amino acid sequence shows that CRY] encodes a 14,561-dalton polypeptide that has 88% amino acid sequence homology to the hamster or human S14 ribosomal protein responsible for emetine resistance and 45% homology to Escherichia coli ribosomal protein Sll. Analysis of the DNA sequences upstream from CRYI revealed the presence of three sequences, HOMOLl (consensus, A/TACATCC/TG/ATA/GCA), RPG (consensus, ACCCA/ GTACATT/CT/A), and a thymine-rich sequence, found upstream of more than 20 other cloned yeast genes encoding components of the translational apparatus. We exploited the ability to assay the expression of CRY) in vivo by using the cryptopleurine resistance phenotype to demonstrate that these three consensus sequences are necessary for the transcription of CRY). We previously showed that the upstream promoter element of the yeast RP39A gene consists of these identical sequence motifs. Therefore, we suggest that these three sequences define a consensus promoter element for the genes encoding the yeast translational apparatus. CRY) is one of several hundred yeast genes, including ribosomal protein genes, whose expression is transiently decreased 10-fold upon heat shock. We found that the HOMOLl and RPG consensus sequences are not necessary for the heat shock response of CRY).The biosynthesis, assembly, and function of ribosomes are complicated interrelated processes essential to the function and growth of all cells. Equimolar quantities of more than 70 different ribosomal proteins synthesized in the cytoplasm and four ribosomal RNAs transcribed in the nucleus are assembled into ribosomes in the nucleolus (16). Ribosome biosynthesis is tightly coordinated with the physiological state of the cells. In the yeast Saccharomyces cerevisiae, the rate of synthesis of ribosomal proteins is coordinately decreased in response to heat shock (31) or upon sporulation (33) and is increased when the yeast is shifted from medium containing ethanol as a carbon source to medium containing glucose as a carbon source (29). To study the coordinate expression of ribosomal genes and the role of individual ribosomal components in yeast ribosome assembly and function, we and other workers have cloned a number of yeast ribosomal protein genes (4, 5, 13-15, 22, 35, 37, 39, 60, 72). With few exceptions, these genes are unlinked, occur in two copies in the genome, and contain a single intervening sequence (1, 4, 5, 13-16, 28, 35, 48, 72, 73).Experiments thus far indicate that a variety of control mechanisms operate to coordinate yeast ribosome biosynthesis. Ribosomal protein mRNA transcription, processing, stability, and translation are regulated, as well as the stability of the ribosomal proteins themselves (11, 12, 18, 19, 29-31,...