Home LiteratureArticle Details
PMID: 9566888 Published · ppublish English Comparative Study Journal Article

Ski6p is a homolog of RNA-processing enzymes that affects translation of non-poly(A) mRNAs and 60S ribosomal subunit biogenesis.

Molecular and cellular biology ·Vol. 18 ·No. 5 ·1998-05-00 ·Pages 2688-96

Benard L, Carroll K, Valle RC, Wickner RB

Abstract

We mapped and cloned SKI6 of Saccharomyces cerevisiae, a gene that represses the copy number of the L-A double-stranded RNA virus, and found that it encodes an essential 246-residue protein with homology to a tRNA-processing enzyme, RNase PH. The ski6-2 mutant expressed electroporated non-poly(A) luciferase mRNAs 8- to 10-fold better than did the isogenic wild type. No effect of ski6-2 on expression of uncapped or normal mRNAs was found. Kinetics of luciferase synthesis and direct measurement of radiolabeled electroporated mRNA indicate that the primary effect of Ski6p was on efficiency of translation rather than on mRNA stability. Both ski6 and ski2 mutants show hypersensitivity to hygromycin, suggesting functional alteration of the translation apparatus. The ski6-2 mutant has normal amounts of 40S and 60S ribosomal subunits but accumulates a 38S particle containing 5'-truncated 25S rRNA but no 5.8S rRNA, apparently an incomplete or degraded 60S subunit. This suggests an abnormality in 60S subunit assembly. The ski6-2 mutation suppresses the poor expression of the poly(A)- viral mRNA in a strain deficient in the 60S ribosomal protein L4. Thus, a ski6 mutation bypasses the requirement of the poly(A) tail for translation, allowing better translation of non-poly(A) mRNA, including the L-A virus mRNA which lacks poly(A). We speculate that the derepressed translation of non-poly(A) mRNAs is due to abnormal (but full-size) 60S subunits.

MeSH Terms
Amino Acid Sequence Exoribonucleases/genetics Exosome Multienzyme Ribonuclease Complex Genes, Lethal Hygromycin B/pharmacology Microbial Sensitivity Tests Molecular Sequence Data Protein Biosynthesis RNA Precursors/metabolism RNA Processing, Post-Transcriptional RNA, Fungal/metabolism RNA, Messenger/metabolism RNA, Ribosomal/metabolism RNA, Transfer/metabolism RNA, Viral/metabolism Ribosomes/metabolism Saccharomyces cerevisiae/genetics,virology Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Suppression, Genetic
Chemicals
RNA Precursors RNA, Fungal RNA, Messenger RNA, Ribosomal RNA, Viral SKI6 protein, S cerevisiae Saccharomyces cerevisiae Proteins Hygromycin B RNA, Transfer ribonuclease PH Exoribonucleases Exosome Multienzyme Ribonuclease Complex
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Benard L
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892-0830, USA.
Carroll K
Valle R C
Wickner R B
References (59)
59 references, click to expand
  1. The 3' end of yeast 5.8S rRNA is generated by an exonuclease processing mechanism.
    Genes Dev. 1996 Feb 15;10(4):502-13 PMID: 8600032
  2. Peptidyl-transferase inhibitors have antiviral properties by altering programmed -1 ribosomal frameshifting efficiencies: development of model systems.
    Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6606-11 PMID: 9192612
  3. Killer systems in Saccharomyces cerevisiae: three distinct modes of exclusion of M2 double-stranded RNA by three species of double-stranded RNA, M1, L-A-E, and L-A-HN.
    Mol Cell Biol. 1983 Apr;3(4):654-61 PMID: 6343841
  4. Chromosomal superkiller mutants of Saccharomyces cerevisiae.
    J Bacteriol. 1978 Dec;136(3):1002-7 PMID: 363683
  5. His-154 is involved in the linkage of the Saccharomyces cerevisiae L-A double-stranded RNA virus Gag protein to the cap structure of mRNAs and is essential for M1 satellite virus expression.
    Mol Cell Biol. 1994 Apr;14(4):2664-74 PMID: 8139566
  6. DRS1 to DRS7, novel genes required for ribosome assembly and function in Saccharomyces cerevisiae.
    Mol Cell Biol. 1993 Dec;13(12):7901-12 PMID: 8247005
  7. Processing of pre-ribosomal RNA in Saccharomyces cerevisiae.
    Yeast. 1995 Dec;11(16):1629-50 PMID: 8720068
  8. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs.
    Genetics. 1993 May;134(1):81-150 PMID: 8514151
  9. RNase PH is essential for tRNA processing and viability in RNase-deficient Escherichia coli cells.
    J Biol Chem. 1992 Aug 15;267(23):16015-8 PMID: 1644789
  10. Plasmids controlled exclusion of the K2 killer double-stranded RNA plasmid of yeast.
    Cell. 1980 Aug;21(1):217-26 PMID: 6996833
  11. mRNA poly(A) tail, a 3' enhancer of translational initiation.
    Mol Cell Biol. 1990 Jul;10(7):3441-55 PMID: 1972543
  12. Genome structure and expression of a defective interfering mutant of the killer virus of yeast.
    Virology. 1984 Aug;137(1):20-31 PMID: 6382788
  13. Synthetic lethality of sep1 (xrn1) ski2 and sep1 (xrn1) ski3 mutants of Saccharomyces cerevisiae is independent of killer virus and suggests a general role for these genes in translation control.
    Mol Cell Biol. 1995 May;15(5):2719-27 PMID: 7739552
  14. Tales of poly(A): a review.
    Gene. 1990 Jul 16;91(2):151-8 PMID: 1976572
  15. Ribosomal frameshifting in yeast viruses.
    Yeast. 1995 Sep 30;11(12):1115-27 PMID: 8619310
  16. "Superkiller" mutations suppress chromosomal mutations affecting double-stranded RNA killer plasmid replication in saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1980 Jan;77(1):527-30 PMID: 6987655
  17. Double-stranded RNA viruses of Saccharomyces cerevisiae.
    Microbiol Rev. 1996 Mar;60(1):250-65 PMID: 8852903
  18. Efficient translation of poly(A)-deficient mRNAs in Saccharomyces cerevisiae.
    Genes Dev. 1994 Nov 1;8(21):2629-40 PMID: 7958921
  19. Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers.
    Yeast. 1990 Sep-Oct;6(5):363-6 PMID: 2220072
  20. Purification and characterization of a Saccharomyces cerevisiae exoribonuclease which yields 5'-mononucleotides by a 5' leads to 3' mode of hydrolysis.
    J Biol Chem. 1980 Apr 10;255(7):3080-5 PMID: 6244307
  21. The coat protein of the yeast double-stranded RNA virus L-A attaches covalently to the cap structure of eukaryotic mRNA.
    Mol Cell Biol. 1992 Aug;12(8):3390-8 PMID: 1630453
  22. RNase PH: an Escherichia coli phosphate-dependent nuclease distinct from polynucleotide phosphorylase.
    Proc Natl Acad Sci U S A. 1988 Jul;85(13):4710-4 PMID: 2455297
  23. Translation and M1 double-stranded RNA propagation: MAK18 = RPL41B and cycloheximide curing.
    J Bacteriol. 1995 May;177(10):2887-91 PMID: 7751301
  24. Mutations in nucleolar proteins lead to nucleolar accumulation of polyA+ RNA in Saccharomyces cerevisiae.
    Mol Biol Cell. 1995 Sep;6(9):1103-10 PMID: 8534909
  25. Yeast virus propagation depends critically on free 60S ribosomal subunit concentration.
    Mol Cell Biol. 1995 May;15(5):2772-81 PMID: 7739558
  26. A translational fidelity mutation in the universally conserved sarcin/ricin domain of 25S yeast ribosomal RNA.
    RNA. 1996 Mar;2(3):254-63 PMID: 8608449
  27. A yeast antiviral protein, SKI8, shares a repeated amino acid sequence pattern with beta-subunits of G proteins and several other proteins.
    Yeast. 1993 Jan;9(1):43-51 PMID: 8442386
  28. The allosuppressor gene SAL4 encodes a protein important for maintaining translational fidelity in Saccharomyces cerevisiae.
    Curr Genet. 1988 Dec;14(6):537-43 PMID: 3072098
  29. Depletion of yeast ribosomal proteins L16 or rp59 disrupts ribosome assembly.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2261-74 PMID: 2277060
  30. A 5'----3' exoribonuclease of Saccharomyces cerevisiae: size and novel substrate specificity.
    Arch Biochem Biophys. 1987 Feb 1;252(2):339-47 PMID: 3545079
  31. A turnover pathway for both stable and unstable mRNAs in yeast: evidence for a requirement for deadenylation.
    Genes Dev. 1993 Aug;7(8):1632-43 PMID: 8393418
  32. Evidence that the SKI antiviral system of Saccharomyces cerevisiae acts by blocking expression of viral mRNA.
    Mol Cell Biol. 1993 Jul;13(7):4331-41 PMID: 8321235
  33. Genetic Control of L-a and L-(Bc) Dsrna Copy Number in Killer Systems of SACCHAROMYCES CEREVISIAE.
    Genetics. 1984 Jun;107(2):199-217 PMID: 17246214
  34. Protein synthesis in yeast. Structural and functional analysis of the gene encoding elongation factor 3.
    J Biol Chem. 1990 Sep 15;265(26):15838-44 PMID: 2203789
  35. The complete sequence of a 9000 bp fragment of the right arm of Saccharomyces cerevisiae chromosome VII contains four previously unknown open reading frames.
    Yeast. 1995 Sep 15;11(11):1087-91 PMID: 7502584
  36. Identification and characterization of a human cDNA homologous to yeast SKI2.
    Genomics. 1995 Feb 10;25(3):660-6 PMID: 7759100
  37. Frameshift suppression in Saccharomyces cerevisiae. IV. New suppressors among spontaneous co-revertants of the Group II his4-206 and leu 2-3 frameshift mutations.
    Genetics. 1982 Jul-Aug;101(3-4):345-67 PMID: 6757051
  38. Ribosome concentration contributes to discrimination against poly(A)- mRNA during translation initiation in Saccharomyces cerevisiae.
    J Biol Chem. 1997 Feb 28;272(9):6004-10 PMID: 9038222
  39. RNA delivery in Saccharomyces cerevisiae using electroporation.
    Yeast. 1992 Dec;8(12):1007-14 PMID: 1284101
  40. Structure and nuclear localization signal of the SKI3 antiviral protein of Saccharomyces cerevisiae.
    Yeast. 1989 May-Jun;5(3):149-58 PMID: 2660461
  41. A -1 ribosomal frameshift in a double-stranded RNA virus of yeast forms a gag-pol fusion protein.
    Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):174-8 PMID: 1986362
  42. Turnover mechanisms of the stable yeast PGK1 mRNA.
    Mol Cell Biol. 1995 Apr;15(4):2145-56 PMID: 7891709
  43. Ribonucleases, tRNA nucleotidyltransferase, and the 3' processing of tRNA.
    Prog Nucleic Acid Res Mol Biol. 1990;39:209-40 PMID: 2247609
  44. Temperature-sensitive mutations demonstrate roles for yeast fibrillarin in pre-rRNA processing, pre-rRNA methylation, and ribosome assembly.
    Cell. 1993 Feb 12;72(3):443-57 PMID: 8431947
  45. Superkiller mutations in Saccharomyces cerevisiae suppress exclusion of M2 double-stranded RNA by L-A-HN and confer cold sensitivity in the presence of M and L-A-HN.
    Mol Cell Biol. 1984 Apr;4(4):761-70 PMID: 6371496
  46. Ribosomal frameshifting efficiency and gag/gag-pol ratio are critical for yeast M1 double-stranded RNA virus propagation.
    J Virol. 1992 Jun;66(6):3669-76 PMID: 1583726
  47. The exosome: a conserved eukaryotic RNA processing complex containing multiple 3'-->5' exoribonucleases.
    Cell. 1997 Nov 14;91(4):457-66 PMID: 9390555
  48. The translation machinery and 70 kd heat shock protein cooperate in protein synthesis.
    Cell. 1992 Oct 2;71(1):97-105 PMID: 1394434
  49. The 5' ends of yeast killer factor RNAs are pppGp.
    Nucleic Acids Res. 1976 Oct;3(10):2427-36 PMID: 792814
  50. NSR1 is required for pre-rRNA processing and for the proper maintenance of steady-state levels of ribosomal subunits.
    Mol Cell Biol. 1992 Sep;12(9):3865-71 PMID: 1508189
  51. Starting at the beginning, middle, and end: translation initiation in eukaryotes.
    Cell. 1997 Jun 13;89(6):831-8 PMID: 9200601
  52. Characterization of Escherichia coli RNase PH.
    J Biol Chem. 1992 Aug 25;267(24):17153-8 PMID: 1512253
  53. Yeast cells lacking 5'-->3' exoribonuclease 1 contain mRNA species that are poly(A) deficient and partially lack the 5' cap structure.
    Mol Cell Biol. 1993 Aug;13(8):4826-35 PMID: 8336719
  54. Direct cloning of yeast genes from an ordered set of lambda clones in Saccharomyces cerevisiae by recombination in vivo.
    Genetics. 1993 May;134(1):151-7 PMID: 8514124
  55. Human helicase gene SKI2W in the HLA class III region exhibits striking structural similarities to the yeast antiviral gene SKI2 and to the human gene KIAA0052: emergence of a new gene family.
    Nucleic Acids Res. 1995 Jun 25;23(12):2120-6 PMID: 7610041
  56. Decoying the cap- mRNA degradation system by a double-stranded RNA virus and poly(A)- mRNA surveillance by a yeast antiviral system.
    Mol Cell Biol. 1995 May;15(5):2763-71 PMID: 7739557
  57. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency.
    Genes Dev. 1991 Nov;5(11):2108-16 PMID: 1682219
  58. 2.2 Mb of contiguous nucleotide sequence from chromosome III of C. elegans.
    Nature. 1994 Mar 3;368(6466):32-8 PMID: 7906398
  59. Mutants of eukaryotic initiation factor eIF-4E with altered mRNA cap binding specificity reprogram mRNA selection by ribosomes in Saccharomyces cerevisiae.
    J Biol Chem. 1996 Mar 22;271(12):7030-7 PMID: 8636134
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.5.2688
Published
1998-05-00
Pages
2688-96
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC110648
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.18.5.2688 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2