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Mol Cell Biol. Oct 1990; 10(10): 5235–5243.
PMCID: PMC361207

Ribosomal protein L30 is dispensable in the yeast Saccharomyces cerevisiae.

Abstract

In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B, which shows strong homology to RPL30A. Use of mRNA as a template for a polymerase chain reaction demonstrated that RPL30B contains an intron in its 5' untranslated region. This intron has an unusual 5' splice site, C/GUAUGU. The genomic copies of RPL30A and RPL30B were disrupted by homologous recombination. Growth rates, primer extension, and two-dimensional ribosomal protein analyses of these disruption mutants suggested that RPL30A is responsible for the majority of L30 production. Surprisingly, meiosis of a diploid strain carrying one disrupted RPL30A and one disrupted RPL30B yielded four viable spores. Ribosomes from haploid cells carrying both disrupted genes had no detectable L30, yet such cells grew with a doubling time only 30% longer than that of wild-type cells. Furthermore, depletion of L30 did not alter the ratio of 60S to 40S ribosomal subunits, suggesting that there is no serious effect on the assembly of 60S subunits. Polysome profiles, however, suggest that the absence of L30 leads to the formation of stalled translation initiation complexes.

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  • Abovich N, Gritz L, Tung L, Rosbash M. Effect of RP51 gene dosage alterations on ribosome synthesis in Saccharomyces cerevisiae. Mol Cell Biol. 1985 Dec;5(12):3429–3435. [PMC free article] [PubMed]
  • Carlson M, Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. [PubMed]
  • Dabbs ER, Ehrlich R, Hasenbank R, Schroeter BH, Stöffler-Meilicke M, Stöffler G. Mutants of Escherichia coli lacking ribosomal protein L1. J Mol Biol. 1981 Jul 15;149(4):553–578. [PubMed]
  • Dabbs ER, Hasenbank R, Kastner B, Rak KH, Wartusch B, Stöffler G. Immunological studies of Escherichia coli mutants lacking one or two ribosomal proteins. Mol Gen Genet. 1983;192(3):301–308. [PubMed]
  • Dabeva MD, Warner JR. The yeast ribosomal protein L32 and its gene. J Biol Chem. 1987 Nov 25;262(33):16055–16059. [PubMed]
  • Elion EA, Warner JR. The major promoter element of rRNA transcription in yeast lies 2 kb upstream. Cell. 1984 Dec;39(3 Pt 2):663–673. [PubMed]
  • Finley D, Bartel B, Varshavsky A. The tails of ubiquitin precursors are ribosomal proteins whose fusion to ubiquitin facilitates ribosome biogenesis. Nature. 1989 Mar 30;338(6214):394–401. [PubMed]
  • Fried HM, Nam HG, Loechel S, Teem J. Characterization of yeast strains with conditionally expressed variants of ribosomal protein genes tcm1 and cyh2. Mol Cell Biol. 1985 Jan;5(1):99–108. [PMC free article] [PubMed]
  • Fried HM, Pearson NJ, Kim CH, Warner JR. The genes for fifteen ribosomal proteins of Saccharomyces cerevisiae. J Biol Chem. 1981 Oct 10;256(19):10176–10183. [PubMed]
  • Grunstein M, Hogness DS. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. [PMC free article] [PubMed]
  • Helser TL, Baan RA, Dahlberg AE. Characterization of a 40S ribosomal subunit complex in polyribosomes of Saccharomyces cerevisiae treated with cycloheximide. Mol Cell Biol. 1981 Jan;1(1):51–57. [PMC free article] [PubMed]
  • Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. [PMC free article] [PubMed]
  • Ju QD, Morrow BE, Warner JR. REB1, a yeast DNA-binding protein with many targets, is essential for growth and bears some resemblance to the oncogene myb. Mol Cell Biol. 1990 Oct;10(10):5226–5234. [PMC free article] [PubMed]
  • Kirsebom LA, Isaksson LA. Involvement of ribosomal protein L7/L12 in control of translational accuracy. Proc Natl Acad Sci U S A. 1985 Feb;82(3):717–721. [PMC free article] [PubMed]
  • Kraft R, Tardiff J, Krauter KS, Leinwand LA. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544–549. [PubMed]
  • Kruse C, Johnson SP, Warner JR. Phosphorylation of the yeast equivalent of ribosomal protein S6 is not essential for growth. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7515–7519. [PMC free article] [PubMed]
  • Langford CJ, Gallwitz D. Evidence for an intron-contained sequence required for the splicing of yeast RNA polymerase II transcripts. Cell. 1983 Jun;33(2):519–527. [PubMed]
  • Lotti M, Dabbs ER, Hasenbank R, Stöffler-Meilicke M, Stöffler G. Characterisation of a mutant from Escherichia coli lacking protein L15 and localisation of protein L15 by immuno-electron microscopy. Mol Gen Genet. 1983;192(3):295–300. [PubMed]
  • Mitra G, Warner JR. A yeast ribosomal protein gene whose intron is in the 5' leader. J Biol Chem. 1984 Jul 25;259(14):9218–9224. [PubMed]
  • Molenaar CM, Woudt LP, Jansen AE, Mager WH, Planta RJ, Donovan DM, Pearson NJ. Structure and organization of two linked ribosomal protein genes in yeast. Nucleic Acids Res. 1984 Oct 11;12(19):7345–7358. [PMC free article] [PubMed]
  • Nieuwint RT, Mager WH, Maurer KC, Planta RJ. Mutational analysis of the upstream activation site of yeast ribosomal protein genes. Curr Genet. 1989 Apr;15(4):247–251. [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]
  • Redman KL, Rechsteiner M. Identification of the long ubiquitin extension as ribosomal protein S27a. Nature. 1989 Mar 30;338(6214):438–440. [PubMed]
  • Remacha M, Sáenz-Robles MT, Vilella MD, Ballesta JP. Independent genes coding for three acidic proteins of the large ribosomal subunit from Saccharomyces cerevisiae. J Biol Chem. 1988 Jul 5;263(19):9094–9101. [PubMed]
  • Rigby PW, Dieckmann M, Rhodes C, Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. [PubMed]
  • Rotenberg MO, Moritz M, Woolford JL., Jr Depletion of Saccharomyces cerevisiae ribosomal protein L16 causes a decrease in 60S ribosomal subunits and formation of half-mer polyribosomes. Genes Dev. 1988 Feb;2(2):160–172. [PubMed]
  • Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. [PubMed]
  • Sachs AB, Davis RW. The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation. Cell. 1989 Sep 8;58(5):857–867. [PubMed]
  • Sachs AB, Davis RW. Translation initiation and ribosomal biogenesis: involvement of a putative rRNA helicase and RPL46. Science. 1990 Mar 2;247(4946):1077–1079. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Stöffler G, Cundliffe E, Stöffler-Meilicke M, Dabbs ER. Mutants of Escherichia coli lacking ribosomal protein L11. J Biol Chem. 1980 Nov 10;255(21):10517–10522. [PubMed]
  • Stöffler G, Noah M, Stöffler-Meilicke M, Dabbs ER. The localization of protein L19 on the surface of 50 S subunits of Escherichia coli aided by the use of mutants lacking protein L19. J Biol Chem. 1984 Apr 10;259(7):4521–4526. [PubMed]
  • Stöffler-Meilicke M, Dabbs ER, Albrecht-Ehrlich R, Stöffler G. A mutant from Escherichia coli which lacks ribosomal proteins S17 and L29 used to localize these two proteins on the ribosomal surface. Eur J Biochem. 1985 Aug 1;150(3):485–490. [PubMed]
  • Subramanian AR, Dabbs ER. Functional studies on ribosomes lacking protein L1 from mutant Escherichia coli. Eur J Biochem. 1980 Nov;112(2):425–430. [PubMed]
  • Teem JL, Abovich N, Kaufer NF, Schwindinger WF, Warner JR, Levy A, Woolford J, Leer RJ, van Raamsdonk-Duin MM, Mager WH, et al. A comparison of yeast ribosomal protein gene DNA sequences. Nucleic Acids Res. 1984 Nov 26;12(22):8295–8312. [PMC free article] [PubMed]
  • Thomas BJ, Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. [PubMed]
  • Warner JR. The assembly of ribosomes in yeast. J Biol Chem. 1971 Jan 25;246(2):447–454. [PubMed]
  • Warner JR. Synthesis of ribosomes in Saccharomyces cerevisiae. Microbiol Rev. 1989 Jun;53(2):256–271. [PMC free article] [PubMed]
  • Warner JR, Gorenstein C. The synthesis of eucaryotic ribosomal proteins in vitro. Cell. 1977 May;11(1):201–212. [PubMed]
  • Warner JR, Gorenstein C. The ribosomal proteins of Saccharomyces cerevisiae. Methods Cell Biol. 1978;20:45–60. [PubMed]
  • Warner JR, Mitra G, Schwindinger WF, Studeny M, Fried HM. Saccharomyces cerevisiae coordinates accumulation of yeast ribosomal proteins by modulating mRNA splicing, translational initiation, and protein turnover. Mol Cell Biol. 1985 Jun;5(6):1512–1521. [PMC free article] [PubMed]
  • Woolford JL., Jr Nuclear pre-mRNA splicing in yeast. Yeast. 1989 Nov-Dec;5(6):439–457. [PubMed]

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