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Mol Cell Biol. 1988 May; 8(5): 2184–2194.
PMCID: PMC363400

Transcription interferes with elements important for chromosome maintenance in Saccharomyces cerevisiae.


Transcription directed into a Saccharomyces cerevisiae autonomously replicating sequence (ARS) causes high-frequency loss of minichromosomes. Conditionally stable artificial yeast chromosomes were constructed that contain an inducible GAL promoter upstream of ARS1. Under growth conditions in which the promoter was inactive, these chromosomes were mitotically stable; however, when the GAL promoter was induced, the chromosomes became extremely unstable as a result of transcriptional impairment of ARS function. This interference by the GAL promoter occurred only in cis but can occur from either side of ARS1. Transcriptional interference of ARS function can be monitored readily by using a visual colony-color assay (P. Hieter, C. Mann, M. Snyder, and R.W. Davis, Cell 40:381-392, 1985), which was further developed as a sensitive in vivo assay for sequences which rescue ARS from transcription. DNA fragments from the 3' ends of genes, inserted downstream of the GAL promoter, protected ARS function from transcriptional interference. This assay is expected to be independent of both RNA transcript stability and processing. Philippsen et al. have shown that transcription into a yeast centromere inhibits CEN function in vivo (L. Panzeri, I. Groth-Clausen, J. Shepard, A. Stotz, and P. Philippsen, Chromosomes Today 8:46-58, 1984). We identified two 200- to 300-base-pair DNA fragments flanking CEN4 that rescued ARS1 from transcription. Both of these fragments protected ARS from transcription when inserted in either orientation. The 3' ends of stable transcripts are encoded by fragments that protected the ARS from transcription, suggesting that the protection was achieved by transcription termination. It is suggested that protection of elements important for the replication and segregation of eucaryotic chromosomes from transcription is necessary for their proper function in vivo.

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  • Bedinger P, Hochstrasser M, Jongeneel CV, Alberts BM. Properties of the T4 bacteriophage DNA replication apparatus: the T4 dda DNA helicase is required to pass a bound RNA polymerase molecule. Cell. 1983 Aug;34(1):115–123. [PubMed]
  • Bloom KS, Carbon J. Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes. Cell. 1982 Jun;29(2):305–317. [PubMed]
  • Bram RJ, Kornberg RD. Isolation of a Saccharomyces cerevisiae centromere DNA-binding protein, its human homolog, and its possible role as a transcription factor. Mol Cell Biol. 1987 Jan;7(1):403–409. [PMC free article] [PubMed]
  • Brewer BJ, Fangman WL. The localization of replication origins on ARS plasmids in S. cerevisiae. Cell. 1987 Nov 6;51(3):463–471. [PubMed]
  • Buchman AR, Kimmerly WJ, Rine J, Kornberg RD. 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):210–225. [PMC free article] [PubMed]
  • Celniker SE, Sweder K, Srienc F, Bailey JE, Campbell JL. Deletion mutations affecting autonomously replicating sequence ARS1 of Saccharomyces cerevisiae. Mol Cell Biol. 1984 Nov;4(11):2455–2466. [PMC free article] [PubMed]
  • Chlebowicz-Sledziewska E, Sledziewski AZ. Construction of multicopy yeast plasmids with regulated centromere function. Gene. 1985;39(1):25–31. [PubMed]
  • Clarke L, Carbon J. Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature. 1980 Oct 9;287(5782):504–509. [PubMed]
  • Dani GM, Zakian VA. Mitotic and meiotic stability of linear plasmids in yeast. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3406–3410. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Fitzgerald-Hayes M, Clarke L, Carbon J. Nucleotide sequence comparisons and functional analysis of yeast centromere DNAs. Cell. 1982 May;29(1):235–244. [PubMed]
  • Futcher B, Carbon J. Toxic effects of excess cloned centromeres. Mol Cell Biol. 1986 Jun;6(6):2213–2222. [PMC free article] [PubMed]
  • Hieter P, Mann C, Snyder M, Davis RW. Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss. Cell. 1985 Feb;40(2):381–392. [PubMed]
  • Hieter P, Pridmore D, Hegemann JH, Thomas M, Davis RW, Philippsen P. Functional selection and analysis of yeast centromeric DNA. Cell. 1985 Oct;42(3):913–921. [PubMed]
  • Hill A, Bloom K. Genetic manipulation of centromere function. Mol Cell Biol. 1987 Jul;7(7):2397–2405. [PMC free article] [PubMed]
  • Huberman JA, Spotila LD, Nawotka KA, el-Assouli SM, Davis LR. The in vivo replication origin of the yeast 2 microns plasmid. Cell. 1987 Nov 6;51(3):473–481. [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]
  • 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]
  • Kearsey S. Structural requirements for the function of a yeast chromosomal replicator. Cell. 1984 May;37(1):299–307. [PubMed]
  • Koshland D, Kent JC, Hartwell LH. Genetic analysis of the mitotic transmission of minichromosomes. Cell. 1985 Feb;40(2):393–403. [PubMed]
  • Lark KG. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. [PubMed]
  • Long CM, Brajkovich CM, Scott JF. Alternative model for chromatin organization of the Saccharomyces cerevisiae chromosomal DNA plasmid TRP1 RI circle (YARp1). Mol Cell Biol. 1985 Nov;5(11):3124–3130. [PMC free article] [PubMed]
  • Mann C, Davis RW. Structure and sequence of the centromeric DNA of chromosome 4 in Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jan;6(1):241–245. [PMC free article] [PubMed]
  • Marczynski GT, Jaehning JA. A transcription map of a yeast centromere plasmid: unexpected transcripts and altered gene expression. Nucleic Acids Res. 1985 Dec 9;13(23):8487–8506. [PMC free article] [PubMed]
  • McLaughlin CS, Warner JR, Edmonds M, Nakazato H, Vaughan MH. Polyadenylic acid sequences in yeast messenger ribonucleic acid. J Biol Chem. 1973 Feb 25;248(4):1466–1471. [PubMed]
  • Murray AW, Szostak JW. Construction of artificial chromosomes in yeast. Nature. 1983 Sep 15;305(5931):189–193. [PubMed]
  • Murray JA, Cesareni G. Functional analysis of the yeast plasmid partition locus STB. EMBO J. 1986 Dec 1;5(12):3391–3399. [PMC free article] [PubMed]
  • Palzkill TG, Oliver SG, Newlon CS. DNA sequence analysis of ARS elements from chromosome III of Saccharomyces cerevisiae: identification of a new conserved sequence. Nucleic Acids Res. 1986 Aug 11;14(15):6247–6264. [PMC free article] [PubMed]
  • Shore D, Nasmyth K. Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements. Cell. 1987 Dec 4;51(5):721–732. [PubMed]
  • Snyder M, Buchman AR, Davis RW. Bent DNA at a yeast autonomously replicating sequence. Nature. 1986 Nov 6;324(6092):87–89. [PubMed]
  • Snyder M, Davidson N. Two gene families clustered in a small region of the Drosophila genome. J Mol Biol. 1983 May 15;166(2):101–118. [PubMed]
  • Sogin SJ, Saunders CA. Fluctuation in polyadenylate size and content in exponential- and stationary-phase cells of Saccharomyces cerevisiae. J Bacteriol. 1980 Oct;144(1):74–81. [PMC free article] [PubMed]
  • Stinchcomb DT, Mann C, Davis RW. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. [PubMed]
  • Stinchcomb DT, Struhl K, Davis RW. Isolation and characterisation of a yeast chromosomal replicator. Nature. 1979 Nov 1;282(5734):39–43. [PubMed]
  • Strich R, Woontner M, Scott JF. Mutations in ARS1 increase the rate of simple loss of plasmids in Saccharomyces cerevisiae. Yeast. 1986 Sep;2(3):169–178. [PubMed]
  • Blackburn EH. The molecular structure of centromeres and telomeres. Annu Rev Biochem. 1984;53:163–194. [PubMed]
  • Szostak JW, Blackburn EH. Cloning yeast telomeres on linear plasmid vectors. Cell. 1982 May;29(1):245–255. [PubMed]
  • Wolffe AP, Brown DD. DNA replication in vitro erases a Xenopus 5S RNA gene transcription complex. Cell. 1986 Oct 24;47(2):217–227. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Yarger JG, Armilei G, Gorman MC. Transcription terminator-like element within a Saccharomyces cerevisiae promoter region. Mol Cell Biol. 1986 Apr;6(4):1095–1101. [PMC free article] [PubMed]
  • Zaret KS, Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. [PubMed]
  • Zaret KS, Sherman F. Mutationally altered 3' ends of yeast CYC1 mRNA affect transcript stability and translational efficiency. J Mol Biol. 1984 Jul 25;177(1):107–135. [PubMed]
  • Zyskind JW, Deen LT, Smith DW. Temporal sequence of events during the initiation process in Escherichia coli deoxyribonucleic acid replication: roles of the dnaA and dnaC gene products and ribonucleic acid polymerase. J Bacteriol. 1977 Mar;129(3):1466–1475. [PMC free article] [PubMed]

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