Home LiteratureArticle Details
PMID: 2697465 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A yeast telomere binding activity binds to two related telomere sequence motifs and is indistinguishable from RAP1.

Current genetics ·Vol. 16 ·No. 4 ·1989-10-00 ·Pages 225-39

Longtine MS, Wilson NM, Petracek ME, Berman J

Abstract

Telomere Binding Activity (TBA), an abundant protein from Saccharomyces cerevisiae, was identified by its ability to bind to telomeric poly(C1-3A) sequence motifs. The substrate specificity of TBA has been analyzed in order to determine whether the activity binds to a unique structure assumed by the irregularly repeating telomeric sequences or whether the activity recognizes and binds to subset of specific sequences found within the telomere repeat tracts. Deletion analysis and DNase I protection assays demonstrate that TBA binds specifically to two poly-(C1-3A) sequences that differ by one nucleotide. The methylation of four guanine residues, located at identical relative positions within these two binding sequences, interferes with TBA binding to the substrates. A synthetic olignucleotide containing a single TBA binding site can function as a TBA binding substrate. The TBA binding site shares homology with the binding sites reported for the Repressor/Activator Protein 1 (RAP1), Translation Upshift Factor (TUF) and General Regulatory Factor (GRFI) transcription factors, and TBA binds directly to RAP1/TUF/GRFI substrate sequences. Yeast TBA preparations and the RAP1 gene product expressed in E. coli cells are both similarly sensitive to in vitro protease digestion. Affinity-purified TBA extracts include a protein indistinguishable from RAP1 in binding specificity, size, and antigenicity. The binding affinity of TBA for the two telomeric poly(C1-3A) binding sites is higher than its affinity for any of the other binding substrates used for its identification. In extracts of yeast spheroplasts prepared by incubation of yeast cells with Zymolyase, an altered, proteolyzed form, of TBA (TBA-S) is present. TBA-S has a faster mobility in gel retardation assays and SDS-PAGE gels, yet it retains the DNA binding properties of standard TBA preparations: it binds to RAP1/TUF/GRFI substrates with the same relative binding affinity and protects poly(C1-3A) tracts from DNase I digestion with a "footprint" identical to that of standard TBA preparations.

MeSH Terms
Base Sequence Binding Sites Binding, Competitive DNA Mutational Analysis DNA, Fungal/genetics DNA-Binding Proteins/genetics,metabolism Electrophoresis, Polyacrylamide Gel Fungal Proteins/genetics,metabolism Immunoblotting Methylation Molecular Sequence Data Plasmids Repetitive Sequences, Nucleic Acid Saccharomyces cerevisiae/genetics Sequence Homology, Nucleic Acid Substrate Specificity Transcription Factors
Chemicals
DNA, Fungal DNA-Binding Proteins Fungal Proteins Transcription Factors
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Longtine M S
Department of Plant Biology, University of Minnesota, St. Paul 55108.
Wilson N M
Petracek M E
Berman J
References (61)
61 references, click to expand
  1. Conserved sequence elements upstream of the gene encoding yeast ribosomal protein L25 are involved in transcription activation.
    EMBO J. 1986 May;5(5):1037-40 PMID: 3013611
  2. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  3. Replication and segregation of plasmids containing cis-acting regulatory sites of silent mating-type genes in Saccharomyces cerevisiae are controlled by the SIR genes.
    Mol Cell Biol. 1987 Dec;7(12):4225-37 PMID: 3325822
  4. A gradient of bicoid protein in Drosophila embryos.
    Cell. 1988 Jul 1;54(1):83-93 PMID: 3383244
  5. Characterization of two telomeric DNA processing reactions in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Nov;8(11):4642-50 PMID: 3062364
  6. A general upstream binding factor for genes of the yeast translational apparatus.
    EMBO J. 1985 Dec 16;4(13A):3539-47 PMID: 3912170
  7. Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements.
    Cell. 1987 Dec 4;51(5):721-32 PMID: 3315231
  8. A yeast silencer contains sequences that can promote autonomous plasmid replication and transcriptional activation.
    Cell. 1987 Dec 4;51(5):709-19 PMID: 3315230
  9. Connections between transcriptional activators, silencers, and telomeres as revealed by functional analysis of a yeast DNA-binding protein.
    Mol Cell Biol. 1988 Dec;8(12):5086-99 PMID: 3072472
  10. Identification of a telomere-binding activity from yeast.
    Proc Natl Acad Sci U S A. 1986 Jun;83(11):3713-7 PMID: 3520552
  11. The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control.
    Mol Gen Genet. 1976 Jan 16;143(2):119-29 PMID: 765749
  12. A mutant with a defect in telomere elongation leads to senescence in yeast.
    Cell. 1989 May 19;57(4):633-43 PMID: 2655926
  13. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts.
    Cell. 1985 Dec;43(2 Pt 1):405-13 PMID: 3907856
  14. O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation.
    Cell. 1988 Oct 7;55(1):125-33 PMID: 3139301
  15. Regulation of mating-type information in yeast. Negative control requiring sequences both 5' and 3' to the regulated region.
    J Mol Biol. 1984 Jul 5;176(3):307-31 PMID: 6379190
  16. Unusual DNA sequences associated with the ends of yeast chromosomes.
    Nature. 1984 Jul 12-18;310(5973):157-60 PMID: 6377091
  17. Transcriptional regulation of ribosomal proteins during a nutritional upshift in Saccharomyces cerevisiae.
    Mol Cell Biol. 1986 Jul;6(7):2429-35 PMID: 3023933
  18. Cloning of random-sequence oligodeoxynucleotides.
    Gene. 1986;44(2-3):177-83 PMID: 3023181
  19. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  20. The UAS of the yeast PGK gene is composed of multiple functional elements.
    Nucleic Acids Res. 1988 Sep 12;16(17):8245-60 PMID: 3047680
  21. The molecular structure of centromeres and telomeres.
    Annu Rev Biochem. 1984;53:163-94 PMID: 6383193
  22. Analysis of upstream activation sites of yeast ribosomal protein genes.
    Nucleic Acids Res. 1987 Aug 11;15(15):6037-48 PMID: 3627978
  23. Telomeric DNA oligonucleotides form novel intramolecular structures containing guanine-guanine base pairs.
    Cell. 1987 Dec 24;51(6):899-908 PMID: 3690664
  24. Affinity purification of sequence-specific DNA binding proteins.
    Proc Natl Acad Sci U S A. 1986 Aug;83(16):5889-93 PMID: 3461465
  25. Identification of an upstream activation site in the pyruvate decarboxylase structural gene (PDC1) of Saccharomyces cerevisiae.
    Curr Genet. 1988 Nov;14(5):405-12 PMID: 3147147
  26. Anti-influenza response achieved by immunization with a synthetic conjugate.
    Proc Natl Acad Sci U S A. 1982 Jan;79(2):569-73 PMID: 6176996
  27. Purification of a yeast protein that binds to origins of DNA replication and a transcriptional silencer.
    Proc Natl Acad Sci U S A. 1988 Apr;85(7):2120-4 PMID: 3281162
  28. Is there left-handed DNA at the ends of yeast chromosomes?
    Nature. 1983 Mar 3;302(5903):84-6 PMID: 6338397
  29. TUF, the yeast DNA-binding factor specific for UASrpg upstream activating sequences: identification of the protein and its DNA-binding domain.
    Proc Natl Acad Sci U S A. 1987 Jun;84(11):3648-52 PMID: 3295867
  30. Activation of yeast RNA polymerase II transcription by a thymidine-rich upstream element in vitro.
    Proc Natl Acad Sci U S A. 1989 Jan;86(2):486-90 PMID: 2643115
  31. Two genes for ribosomal protein 51 of Saccharomyces cerevisiae complement and contribute to the ribosomes.
    Mol Cell Biol. 1984 Sep;4(9):1871-9 PMID: 6092944
  32. A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis.
    Nature. 1989 Jan 26;337(6205):331-7 PMID: 2463488
  33. Several distinct "CCAAT" box binding proteins coexist in eukaryotic cells.
    Proc Natl Acad Sci U S A. 1988 Feb;85(3):757-61 PMID: 3422457
  34. An agarose gel electrophoresis assay for the detection of DNA-binding activities in yeast cell extracts.
    Methods Enzymol. 1987;155:528-37 PMID: 3323822
  35. Tripartite upstream promoter element essential for expression of Saccharomyces cerevisiae ribosomal protein genes.
    Mol Cell Biol. 1986 Feb;6(2):674-87 PMID: 3023862
  36. Yeast transformation: a model system for the study of recombination.
    Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354-8 PMID: 6273866
  37. Generation of telomere-length heterogeneity in Saccharomyces cerevisiae.
    Proc Natl Acad Sci U S A. 1988 Jan;85(2):534-8 PMID: 3277178
  38. DNA sequences of telomeres maintained in yeast.
    Nature. 1984 Jul 12-18;310(5973):154-7 PMID: 6330571
  39. Roles of two DNA-binding factors in replication, segregation and transcriptional repression mediated by a yeast silencer.
    EMBO J. 1988 Jul;7(7):2241-53 PMID: 3046937
  40. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  41. A consensus sequence polymer inhibits in vivo expression of heat shock genes.
    Mol Cell Biol. 1986 Sep;6(9):3200-6 PMID: 3023967
  42. Origin of concatemeric T7 DNA.
    Nat New Biol. 1972 Oct 18;239(94):197-201 PMID: 4507727
  43. The telomere terminal transferase of Tetrahymena is a ribonucleoprotein enzyme with two kinds of primer specificity.
    Cell. 1987 Dec 24;51(6):887-98 PMID: 3319189
  44. In situ detection of sequence-specific DNA binding activity specified by a recombinant bacteriophage.
    Genes Dev. 1988 Jul;2(7):801-6 PMID: 3061875
  45. Identification of silencer binding proteins from yeast: possible roles in SIR control and DNA replication.
    EMBO J. 1987 Feb;6(2):461-7 PMID: 15981337
  46. Telomere terminal transferase activity in the hypotrichous ciliate Oxytricha nova and a model for replication of the ends of linear DNA molecules.
    Nucleic Acids Res. 1988 Jul 25;16(14B):6953-72 PMID: 3136437
  47. Identification of yeast mutants with altered telomere structure.
    Proc Natl Acad Sci U S A. 1986 Mar;83(5):1398-402 PMID: 3513174
  48. Cloning yeast telomeres on linear plasmid vectors.
    Cell. 1982 May;29(1):245-55 PMID: 6286143
  49. Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Jan;8(1):210-25 PMID: 3275867
  50. The Saccharomyces cerevisiae chromosome III left telomere has a type X, but not a type Y', ARS region.
    Mol Cell Biol. 1986 Apr;6(4):1352-6 PMID: 3537702
  51. Protein blotting: principles and applications.
    Anal Biochem. 1983 May;131(1):1-15 PMID: 6193725
  52. The Fusion of Broken Ends of Chromosomes Following Nuclear Fusion.
    Proc Natl Acad Sci U S A. 1942 Nov;28(11):458-63 PMID: 16578057
  53. A multiplicity of CCAAT box-binding proteins.
    Cell. 1987 Sep 11;50(6):863-72 PMID: 3476205
  54. The Stability of Broken Ends of Chromosomes in Zea Mays.
    Genetics. 1941 Mar;26(2):234-82 PMID: 17247004
  55. Purification of nuclear factor I by DNA recognition site affinity chromatography.
    J Biol Chem. 1986 Jan 25;261(3):1398-408 PMID: 3003068
  56. Transcriptional control of yeast ribosomal protein synthesis during carbon-source upshift.
    Nucleic Acids Res. 1987 Dec 23;15(24):10133-44 PMID: 3320961
  57. CDC17: an essential gene that prevents telomere elongation in yeast.
    Cell. 1985 Aug;42(1):249-57 PMID: 3893744
  58. Elaboration of telomeres in yeast: recognition and modification of termini from Oxytricha macronuclear DNA.
    Proc Natl Acad Sci U S A. 1984 Mar;81(5):1475-9 PMID: 6324194
  59. Distribution of telomere-associated sequences on natural chromosomes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 May;8(5):2257-60 PMID: 3290654
  60. Production of single-stranded plasmid DNA.
    Methods Enzymol. 1987;153:3-11 PMID: 3323803
  61. Organization of DNA sequences and replication origins at yeast telomeres.
    Cell. 1983 Jun;33(2):563-73 PMID: 6345000
Article Info
Journal
Current genetics
Abbr.
Curr Genet
ISSN
0172-8083
DOI
10.1007/BF00422108
Published
1989-10-00
Pages
225-39
Language
English
Region
United States
NLM ID
8004904
Subset
IM
Grants
NIGMS NIH HHS · GM38626 · United States
External Links
PubMed source DOI: 10.1007/BF00422108 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: [email protected]

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