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PMID: 1922070 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Assembly of 60S ribosomal subunits is perturbed in temperature-sensitive yeast mutants defective in ribosomal protein L16.

Molecular and cellular biology ·Vol. 11 ·No. 11 ·1991-11-00 ·Pages 5681-92

Moritz M, Pulaski BA, Woolford JL

Abstract

Temperature-sensitive mutants defective in 60S ribosomal subunit protein L16 of Saccharomyces cerevisiae were isolated through hydroxylamine mutagenesis of the RPL16B gene and plasmid shuffling. Two heat-sensitive and two cold-sensitive isolates were characterized. The growth of the four mutants is inhibited at their restrictive temperatures. However, many of the cells remain viable if returned to their permissive temperatures. All of the mutants are deficient in 60S ribosomal subunits and therefore accumulate translational preinitiation complexes. Three of the mutants exhibit a shortage of mature 25S rRNA, and one accumulates rRNA precursors. The accumulation of rRNA precursors suggests that ribosome assembly may be slowed in this mutant. These phenotypes lead us to propose that mutants containing the rpl16b alleles are defective for 60S subunit assembly rather than function. In the mutant carrying the rpl16b-1 allele, ribosomes initiate translation at the noncanonical codon AUA, at least on the rpl16b-1 mRNA, bringing to light a possible connection between the rate and the fidelity of translation initiation.

Related Genes
rpl16b-1
MeSH Terms
Amino Acid Sequence Animals Codon Fungal Proteins/analysis Genes, Fungal Genotype Hydroxylamine Hydroxylamines/pharmacology Molecular Sequence Data Mutagenesis Polyribosomes/metabolism RNA Precursors/genetics,metabolism RNA, Ribosomal/genetics Ribosomal Proteins/analysis,genetics,metabolism Ribosomes/metabolism Saccharomyces cerevisiae/drug effects,genetics,metabolism Sequence Homology, Nucleic Acid Temperature
Chemicals
Codon Fungal Proteins Hydroxylamines RNA Precursors RNA, Ribosomal Ribosomal Proteins ribosomal protein L16 Hydroxylamine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Moritz M
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Pulaski B A
Woolford J L
References (66)
66 references, click to expand
  1. Sequence and structural features associated with translational initiator regions in yeast--a review.
    Gene. 1987;59(1):1-18 PMID: 3325335
  2. Characterization of a 40S ribosomal subunit complex in polyribosomes of Saccharomyces cerevisiae treated with cycloheximide.
    Mol Cell Biol. 1981 Jan;1(1):51-7 PMID: 6765595
  3. Saccharomyces cerevisiae coordinates accumulation of yeast ribosomal proteins by modulating mRNA splicing, translational initiation, and protein turnover.
    Mol Cell Biol. 1985 Jun;5(6):1512-21 PMID: 3897837
  4. Cold-sensitive mutations in Salmonella typhimurium which affect ribosome synthesis.
    J Bacteriol. 1969 Mar;97(3):1298-304 PMID: 4887510
  5. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast.
    Cell. 1987 Mar 27;48(6):1047-60 PMID: 3030557
  6. Further characterization of recessive suppression in yeast. Isolation of the low-temperature sensitive mutant of Saccharomyces cerevisiae defective in the assembly of 60 S ribosomal subunit.
    Biochim Biophys Acta. 1981 Jun 26;654(1):149-55 PMID: 7023541
  7. Mutation in Saccharomyces cerevisiae conferring streptomycin and cold sensitivity by affecting ribosome formation and function.
    J Bacteriol. 1974 May;118(2):319-28 PMID: 4133351
  8. The complete amino acid sequences of the 5 S rRNA binding proteins L5 and L18 from the moderate thermophile Bacillus stearothermophilus ribosome.
    FEBS Lett. 1987 Jan 1;210(1):85-90 PMID: 3542562
  9. Suppressors of temperature-sensitive mutations in a ribosomal protein gene, rpsL (S12), of Escherichia coli K12.
    Mol Gen Genet. 1985;199(3):381-7 PMID: 3162077
  10. Temperature-sensitive mutants with alterations in ribosomal protein L24 and isolation of intra- and extragenic suppressor mutants.
    Methods Enzymol. 1988;164:706-9 PMID: 3241553
  11. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  12. Depletion of U14 small nuclear RNA (snR128) disrupts production of 18S rRNA in Saccharomyces cerevisiae.
    Mol Cell Biol. 1990 Mar;10(3):1145-52 PMID: 2406561
  13. Downstream secondary structure facilitates recognition of initiator codons by eukaryotic ribosomes.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8301-5 PMID: 2236042
  14. The primary structure of the gene encoding yeast ribosomal protein L16.
    FEBS Lett. 1984 Oct 1;175(2):371-6 PMID: 6090215
  15. Cloning and analysis of the spc ribosomal protein operon of Bacillus subtilis: comparison with the spc operon of Escherichia coli.
    Nucleic Acids Res. 1989 Sep 25;17(18):7469-86 PMID: 2508062
  16. Improved tools for biological sequence comparison.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8 PMID: 3162770
  17. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli.
    Gene. 1987;57(2-3):267-72 PMID: 3319781
  18. Molecular characterization of the yeast PRT1 gene in which mutations affect translation initiation and regulation of cell proliferation.
    J Biol Chem. 1987 Feb 25;262(6):2845-51 PMID: 3029094
  19. Coordinate control of syntheses of ribosomal ribonucleic acid and ribosomal proteins during nutritional shift-up in Saccharomyces cerevisiae.
    Mol Cell Biol. 1981 Nov;1(11):1007-15 PMID: 7050661
  20. Genetic selection for mutations that reduce or abolish ribosomal recognition of the HIS4 translational initiator region.
    Mol Cell Biol. 1988 Jul;8(7):2955-63 PMID: 3043200
  21. The 5-S RNA binding protein from yeast (Saccharomyces cerevisiae) ribosomes. Evolution of the eukaryotic 5-S RNA binding protein.
    Eur J Biochem. 1979 Dec 17;102(2):573-82 PMID: 393511
  22. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase.
    Biotechniques. 1988 Jun;6(6):544-6, 549 PMID: 3273187
  23. RNA-protein interactions in 30S ribosomal subunits: folding and function of 16S rRNA.
    Science. 1989 May 19;244(4906):783-90 PMID: 2658053
  24. Toxic effects of excess cloned centromeres.
    Mol Cell Biol. 1986 Jun;6(6):2213-22 PMID: 3537715
  25. Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants.
    Genetics. 1973 Jun;74(2):267-86 PMID: 17248617
  26. A mutation allowing an mRNA secondary structure diminishes translation of Saccharomyces cerevisiae iso-1-cytochrome c.
    Mol Cell Biol. 1985 Aug;5(8):1839-46 PMID: 3018530
  27. Isolation of active ribosomal subunits from yeast.
    Eur J Biochem. 1972 Oct 17;30(1):15-25 PMID: 5086603
  28. Temperature sensitive mutations affecting ribosome synthesis in Saccharomyces cerevisiae.
    J Mol Biol. 1972 Mar 28;65(2):243-57 PMID: 4557193
  29. Functional substitution of mouse ribosomal protein L27' for yeast ribosomal protein L29 in yeast ribosomes.
    Proc Natl Acad Sci U S A. 1989 Jan;86(1):217-21 PMID: 2643099
  30. A trichodermin-resistant mutant of Saccharomyces cerevisiae with an abnormal distribution of native ribosomal subunits.
    Eur J Biochem. 1980;107(1):173-83 PMID: 6995112
  31. Translation initiation and ribosomal biogenesis: involvement of a putative rRNA helicase and RPL46.
    Science. 1990 Mar 2;247(4946):1077-9 PMID: 2408148
  32. A cold-sensitive mutant of Saccharomyces cerevisiae defective in ribosome processing.
    Mol Gen Genet. 1979 Oct 1;175(3):313-23 PMID: 392231
  33. Saccharomyces cerevisiae ribosomes recognize non-AUG initiation codons.
    Mol Cell Biol. 1984 Jul;4(7):1191-7 PMID: 6390186
  34. 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):160-72 PMID: 3282992
  35. Ribosomal RNA synthesis in Saccharomyces cerevisiae.
    J Mol Biol. 1972 Mar 28;65(2):227-42 PMID: 4557192
  36. On the control of ribosomal protein biosynthesis in E. coli. IV. Studies on a temperature-sensitive mutant defective in the assembly of 50S subunits.
    Mol Gen Genet. 1979 Mar 27;171(3):317-25 PMID: 377017
  37. The ribosomal protein gene cluster of Mycoplasma capricolum.
    Mol Gen Genet. 1987 Dec;210(2):314-22 PMID: 3481422
  38. Depletion of yeast ribosomal proteins L16 or rp59 disrupts ribosome assembly.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2261-74 PMID: 2277060
  39. The spc ribosomal protein operon of Escherichia coli: sequence and cotranscription of the ribosomal protein genes and a protein export gene.
    Nucleic Acids Res. 1983 May 11;11(9):2599-616 PMID: 6222285
  40. Conditional expression of RPA190, the gene encoding the largest subunit of yeast RNA polymerase I: effects of decreased rRNA synthesis on ribosomal protein synthesis.
    Mol Cell Biol. 1990 May;10(5):2049-59 PMID: 2183018
  41. The poly(A) binding protein is required for poly(A) shortening and 60S ribosomal subunit-dependent translation initiation.
    Cell. 1989 Sep 8;58(5):857-67 PMID: 2673535
  42. Structure and function of E. coli ribosomes. 8. Cold-sensitive mutants defective in ribosome assembly.
    Proc Natl Acad Sci U S A. 1969 Jun;63(2):384-91 PMID: 4895536
  43. On the control of ribosomal protein biosynthesis in E. coli. III. Studies on a temperature-sensitive mutant defective in the assembly of 30S subunits.
    Mol Gen Genet. 1979 Mar 27;171(3):307-16 PMID: 377016
  44. Genetic analysis of mutations affecting growth of Saccharomyces cerevisiae at low temperature.
    Genetics. 1974 Aug;77(4):651-9 PMID: 4371644
  45. Synthesis of ribosomes in Saccharomyces cerevisiae.
    Microbiol Rev. 1989 Jun;53(2):256-71 PMID: 2666845
  46. A comprehensive set of sequence analysis programs for the VAX.
    Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
  47. A yeast mutant defective in the processing of 27S r-RNA precursor.
    Mol Gen Genet. 1976 Feb 27;144(1):29-37 PMID: 772413
  48. RRP1, a Saccharomyces cerevisiae gene affecting rRNA processing and production of mature ribosomal subunits.
    J Bacteriol. 1987 Apr;169(4):1571-8 PMID: 3549696
  49. Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae.
    Microbiol Rev. 1988 Jun;52(2):248-73 PMID: 3045517
  50. Organization and structure of the Methanococcus transcriptional unit homologous to the Escherichia coli "spectinomycin operon". Implications for the evolutionary relationship of 70 S and 80 S ribosomes.
    J Mol Biol. 1989 Sep 5;209(1):21-36 PMID: 2530355
  51. Structure and expression of the Saccharomyces cerevisiae CRY1 gene: a highly conserved ribosomal protein gene.
    Mol Cell Biol. 1987 May;7(5):1764-75 PMID: 3037334
  52. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics.
    Methods Enzymol. 1987;154:164-75 PMID: 3323810
  53. Characterization of nuclear localizing sequences derived from yeast ribosomal protein L29.
    EMBO J. 1990 Jan;9(1):91-9 PMID: 2104804
  54. The accumulation of three yeast ribosomal proteins under conditions of excess mRNA is determined primarily by fast protein decay.
    Mol Cell Biol. 1988 Jan;8(1):169-75 PMID: 3275866
  55. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs.
    Nucleic Acids Res. 1987 Oct 26;15(20):8125-48 PMID: 3313277
  56. Ribosomal protein synthesis is not regulated at the translational level in Saccharomyces cerevisiae: balanced accumulation of ribosomal proteins L16 and rp59 is mediated by turnover of excess protein.
    Genes Dev. 1988 Jun;2(6):664-76 PMID: 3047007
  57. Structure and function of bacterial ribosomes. XI. Dependence of 50S ribosomal assembly on simultaneous assembly of 30S subunits.
    Proc Natl Acad Sci U S A. 1970 Nov;67(3):1440-7 PMID: 4922290
  58. Isolation of cloned DNA sequences containing ribosomal protein genes from Saccharomyces cerevisiae.
    Cell. 1979 Dec;18(4):1247-59 PMID: 391408
  59. The assembly of ribosomes.
    J Cell Physiol. 1969 Oct;74(2):Suppl 1:241+ PMID: 4902821
  60. Effects of progressive depletion of TCM1 or CYH2 mRNA on Saccharomyces cerevisiae ribosomal protein accumulation.
    Mol Cell Biol. 1986 May;6(5):1535-44 PMID: 3537704
  61. The small nucleolar RNP protein NOP1 (fibrillarin) is required for pre-rRNA processing in yeast.
    EMBO J. 1991 Mar;10(3):573-83 PMID: 1825809
  62. A temperature-sensitive mutation affecting 28S ribosomal RNA production in mammalian cells.
    Proc Natl Acad Sci U S A. 1973 Apr;70(4):1273-7 PMID: 4515626
  63. Genetic analysis of cold-sensitive ribosome maturation mutants of Escherichia coli.
    J Bacteriol. 1974 Mar;117(3):1082-92 PMID: 4591943
  64. Analysis of temperature-sensitive mutant ts 187 of Saccharomyces cerevisiae altered in a component required for the initiation of protein synthesis.
    J Biol Chem. 1982 Sep 25;257(18):10846-51 PMID: 7050121
  65. Ribosome structure, maturation of ribosomal RNA and drug sensitivity in temperature-sensitive mutants of Saccharomyces cerevisiae.
    Mol Gen Genet. 1982;188(3):384-91 PMID: 6761547
  66. The cellular level of yeast ribosomal protein L25 is controlled principally by rapid degradation of excess protein.
    Curr Genet. 1986;10(10):733-9 PMID: 3329033
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.11.5681-5692.1991
Published
1991-11-00
Pages
5681-92
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361939
Subset
IM
Grants
NCI NIH HHS · CA-01000 · United States
NIGMS NIH HHS · GM-28301 · United States
NIGMS NIH HHS · GM08067 · United States
External Links
PubMed source DOI: 10.1128/mcb.11.11.5681-5692.1991 PMC Full Text View journal details
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