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Mol Cell Biol. 1985 January; 5(1): 99–108.
PMCID: PMC366683

Characterization of yeast strains with conditionally expressed variants of ribosomal protein genes tcm1 and cyh2.

Abstract

We placed a regulatory sequence derived from the GAL10 locus of Saccharomyces cerevisiae at various distances from the start sites of transcription of two yeast ribosomal protein genes, tcm1 and cyh2. The hybrid ribosomal protein genes were transcribed at wild-type levels in the presence of galactose. In the absence of galactose, the hybrid genes were transcribed either at a reduced level or essentially not at all. Yeast cells which transcribe the ribosomal protein genes at a reduced rate continued to grow, suggesting that enhanced translation of the ribosomal protein mRNA may permit an adequate rate of synthesis of the corresponding protein. Consistent with this suggestion is the finding that preexisting mRNA decayed at a reduced rate when transcription was halted abruptly by removal of galactose. Yeast cells unable to transcribe tcm1 or cyh2 without galactose did not grow. These conditional lethal strains demonstrate that the ribosomal proteins encoded by tcm1 and cyh2 are essential; furthermore, these strains are potentially useful for isolating mutations in the tcm1 and cyh2 proteins affecting their transport, assembly, or function.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Fried HM, Warner JR. Cloning of yeast gene for trichodermin resistance and ribosomal protein L3. Proc Natl Acad Sci U S A. 1981 Jan;78(1):238–242. [PubMed]
  • Fried HM, Warner JR. Molecular cloning and analysis of yeast gene for cycloheximide resistance and ribosomal protein L29. Nucleic Acids Res. 1982 May 25;10(10):3133–3148. [PMC free article] [PubMed]
  • Grant PG, Schindler D, Davies JE. Mapping of trichodermin resistance in Saccharomyces cerevisiae: a genetic locus for a component of the 60S ribsomal subunit. Genetics. 1976 Aug;83(4):667–673. [PubMed]
  • Guarente L, Yocum RR, Gifford P. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410–7414. [PubMed]
  • Hereford LM, Rosbash M. Regulation of a set of abundant mRNA sequences. Cell. 1977 Mar;10(3):463–467. [PubMed]
  • Hinnen A, Hicks JB, Fink GR. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. [PubMed]
  • Hopper JE, Broach JR, Rowe LB. Regulation of the galactose pathway in Saccharomyces cerevisiae: induction of uridyl transferase mRNA and dependency on GAL4 gene function. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2878–2882. [PubMed]
  • Hu N, Messing J. The making of strand-specific M13 probes. Gene. 1982 Mar;17(3):271–277. [PubMed]
  • Johnston M, Davis RW. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. [PMC free article] [PubMed]
  • Käufer NF, Fried HM, Schwindinger WF, Jasin M, Warner JR. Cycloheximide resistance in yeast: the gene and its protein. Nucleic Acids Res. 1983 May 25;11(10):3123–3135. [PMC free article] [PubMed]
  • Kim CH, Warner JR. Messenger RNA for ribosomal proteins in yeast. J Mol Biol. 1983 Mar 25;165(1):79–89. [PubMed]
  • Messing J, Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. [PubMed]
  • Ng R, Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. [PubMed]
  • Pearson NJ, Fried HM, Warner JR. Yeast use translational control to compensate for extra copies of a ribosomal protein gene. Cell. 1982 Jun;29(2):347–355. [PubMed]
  • Penman S, Fulton A, Capco D, Ben Ze'ev A, Wittelsberger S, Tse CF. Cytoplasmic and nuclear architecture in cells and tissue: form, functions, and mode of assembly. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):1013–1028. [PubMed]
  • Perlman D, Hopper JE. Constitutive synthesis of the GAL4 protein, a galactose pathway regulator in Saccharomyces cerevisiae. Cell. 1979 Jan;16(1):89–95. [PubMed]
  • Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. [PubMed]
  • Schultz LD, Friesen JD. Nucleotide sequence of the tcml gene (ribosomal protein L3) of Saccharomyces cerevisiae. J Bacteriol. 1983 Jul;155(1):8–14. [PMC free article] [PubMed]
  • St John TP, Davis RW. Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridization. Cell. 1979 Feb;16(2):443–452. [PubMed]
  • St John TP, Davis RW. The organization and transcription of the galactose gene cluster of Saccharomyces. J Mol Biol. 1981 Oct 25;152(2):285–315. [PubMed]
  • Teem JL, Rosbash M. Expression of a beta-galactosidase gene containing the ribosomal protein 51 intron is sensitive to the rna2 mutation of yeast. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4403–4407. [PubMed]
  • Thomas PS. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. [PubMed]
  • Warner JR, Gorenstein C. The synthesis of eucaryotic ribosomal proteins in vitro. Cell. 1977 May;11(1):201–212. [PubMed]

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