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

Abstract

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,...

Depletion of Saccharomyces cerevisiae ribosomal protein L16 causes a decrease in 60S ribosomal subunits and formation of half-mer polyribosomes.

“…Levy, in prep.). Although the arrangement of the tripartite consensus promoter motifs common to yeast ribosomal protein genes (Rotenberg and Woolford 1986;Larkin et al 1987) differs between the two RPL16 genes (see Fig. 7; Rotenberg and Woolford 1986), we have not been able to correlate these specific arrangements with levels of transcription of each gene.…”

The organization and expression of a maize ribosomal protein gene family.

“…The 40S subunit ribosomal protein S 14, encoded in yeast by the CRY1 gene, has been highly conserved throughout eukaryotic evolution (Chen et al 1986;Larkin et al 1987). Therefore, this yeast gene seemed likely to be an effective hybridization probe for the isolation of a maize ribosomal protein eDNA.…”

“…When the amino acid alignment shown in Figure 2B is used and excluding gaps are introduced to maintain sequence alignment, 77% of the amino acids (105 of 137) are identical between MCH1 and the yeast S14 ribosomal protein (Larkin et al 1987), and 88% (129 of 147) of the amino acids are identical between MGH1 and the CHO cell S14 ribosomal protein (Rhoads and Roufa 1985). The predicted MCH2 gene product is slightly less conserved relative to the CHO cell and yeast proteins (76 and 87% amino acid identity, respectively).…”

“…The sequence of this ribosomal protein is highly conserved among yeast, mammals, and maize (Chen et al 1986;Larkin et al 1987; this study). The maize S14 ribosomal protein is encoded by a small multigene family with a minimum of three to six members, at least three of which are expressed.…”

Giant panda ribosomal protein S14: cDNA, genomic sequence cloning, sequence analysis, and overexpression

“…The small subunit protein RPS14, the yeast homolog of the bacterial S11 protein, directly binds helix 23 of 18S rRNA (Fewell and Woolford Jr., 1999;Mayolo and Woolford Jr., 2003). Recently, it was reported that RPS14 is an essential protein necessary for the maturation of the 40S ribosomal subunit (Larkin et al, 1987;Moritz et al, 1990;Paulovich et al, 1993;Ford et al, 1999;Jakovljevic et al, 2004) and the processing of 18S pre-rRNA (Ferreira-Cerca et al, 2005;Ebert et al, 2008). And in the 40S subunit, RPS14 helps to form the mRNA exit channel along with its interacting protein RPS5 (Yusupov et al, 2001).…”