Skip to main content
SpringerLink
Log in

The chromatin of theSaccharomyces cerevisiae centromere shows cell-type specific changes

  • Original Articles
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

We have analysed the centromeric chromatin from chromosome XIV ofSaccharomyces cerevisiae at different stages of mitosis with the help of mutants of the cell division cycle. The pattern of centromeric chromatin in cells arrested usingcdc20-1, tub2-401 andcdc15-1 alleles was indistinguishable from that of vegetatively growing cells, indicating that the centromeric complex is constitutively present during mitosis and posably throughout the entire cell cycle. In contrast chromatin isolated from G0 cells and spores exhibited distinct differences in centromeric chromatin probably due to structural rearrangements of the centromeric complex. In particular the alterations found in spores are indicative of an inactive centromeric complex. The differences in centromeric chromatin in spores do not reflect a general reorganisation of the chromatin in this cell type, as the chromatin structure of thePHO3/PHO5 locus in spores was found to be identical to that in vegetative cells under repressed conditions. Thus the structural analysis of the centromere in different cell types provides evidence about the requirement ofCEN DNA/protein complexes in different cell types and in different stages of the cell cycle.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Almer A, Hörz W (1986) Nuclease hypersensitive regions with adjacent positioned nucleosomes mark the gene boundaries of thePHO5/PHO3 locus in yeast. EMBO J 5:2681–2687

    Google Scholar 

  • Baker RE, Masison DC (1990) Isolation of the gene encoding theSaccharomyces cerevisiae centromere-binding protein CP1. Mol Cell Biol 10:2458–2467

    Google Scholar 

  • Baker RE, Fitzgerald-Hayes M, O'Brien TC (1989) Purification of the yeast centromere binding protein CP1 and a mutational analysis of its binding site. J Biol Chem 246:10843–10850

    Google Scholar 

  • Bloom KS, Carbon J (1982) Yeast centromeric DNA is in a highly ordered structure in chromosomes and small circular minichromosomes. Cell 29:305–317

    Google Scholar 

  • Bloom KS, Amaya E, Carbon J, Clarke L, Hill A, Yeh E (1984) Chromatin conformation of yeast centromeres. J Cell Biol 99: 1559–1568

    Google Scholar 

  • Bloom K, Hill A, Kenna M, Saunders M (1989) The structure of a primitive kinetochore. Trends Biochem Sci 14:223–227

    Google Scholar 

  • Bram RJ, Kornberg RD (1987) Isolation of aSaccharomyces cerevisiae centromeric DNA-binding protein, its human homolog, and its possible role as a transcription factor. Mol Cell Biol 7: 403–409

    Google Scholar 

  • Brinkley BR, Ouspenski I, Zinkowski RP (1992) Structure and molecular organization of the centromere-kinetochore complex. Trends Cell Biol 2:15–21

    Google Scholar 

  • Brown JA, Holmes SG, Smith MM (1991) The chromatin structure ofSaccharomyces cerevisiae autonomously replicating sequences changes during the cell division cycle. Mol Cell Biol 11:5301–5311

    Google Scholar 

  • Byers B (1981) In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeastSaccharomyces: life cycle and inheritance. Cold Spring Harbor Laboratory Press, New York, pp 59–96

    Google Scholar 

  • Byers B, Goetsch L (1975) Behavior of spindles and spindle plaques in the cell cycle and conjugation ofSaccharomyces cerevisiae. J Bacteriol 124:511–523

    Google Scholar 

  • Cai M, Davis RW (1989) Purification of a yeast centromere-binding protein that is able to distinguish single base-pair mutations in its recognition site. Mol Cell Biol 9:2544–2550

    Google Scholar 

  • Cai M, Davis RW (1990) Yeast centromere binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy. Cell 61:437–446

    Google Scholar 

  • Cao L, Alani E, Kleckner N (1990) A pathway for generation and processing of double-strand breaks during meiotic recombination inS. cerevisiae. Cell 61:1089–1101

    Google Scholar 

  • Cottarel G, Shero JH, Hieter P, Hegemann JH (1989) A 125-base pairCEN6 DNA fragment is sufficient for complete meiotic and mitotic centromere functions inSaccharomyces cerevisiae. Mol Cell Biol 9:3342–3349

    Google Scholar 

  • Dawes IW, Hardie ID (1974) Selective killing of vegetative cells in sporulated yeast cultures by exposure to diethylether. Mol Gen Genet 131:281–289

    Google Scholar 

  • Diffley JFX, Cocker JH, Dowell SJ, Rowley A (1994) Two steps in the assembly of complexes at yeast replication origins in vivo. Cell 78:303–316

    Google Scholar 

  • Doheny KF, Sorger PK, Hyman AA, Tugendreich S, Spencer F, Hieter P (1993) Identification of essential components of theSaccharomyces cerevisiae kinetochore. Cell 73:761–774

    Google Scholar 

  • Drebot MA, Barnes CA, Singer RA, GC Johnston (1990) Genetic assessment of stationary phase for cells of the yeastSaccharomyces cerevisiae. J Bacteriol 172:3584–3589

    Google Scholar 

  • Esposito RE, Dressler M, Breitenbach M (1991) Identifiying sporulation genes, visualizing synaptonemal complexes, and large-scale spore and spore wall purification. Methods Enzymol 194:110–131

    Google Scholar 

  • Fitzgerald-Hayes M, Clarke L, Carbon J (1982) Nucleotide sequence comparisons and functional analysis of yeast centromeric DNAs. Cell 29:235–244

    Google Scholar 

  • Funabiki H, Hagan I, Uzawa S, Yanagida M (1993) Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast. J Cell Biol 121:961–976

    Google Scholar 

  • Funk M (1990) Analysis of structural determinants of the centromeric chromatin fromSaccharomyces cerevisiae. Ph.D thesis, University of Giessen, Giessen

  • Funk M, Hegemann JH, Philippsen P (1989) Chromatin digestion with restriction endonucleases reveals 150–160 bp of protected DNA in the centromere of chromosome 14 inSaccharomyces cerevisiae. Mol Gen Genet 219:153–160

    Google Scholar 

  • Greenfelder SA, Newlon CS (1992) Replication forks pause at yeast centromeres Mol Cell Biol 12:4056–4066

    Google Scholar 

  • Guacci V, Hogan E, Koshland D (1994) Chromosome condensation and sister chromatid pairing in budding yeast. J Cell Biol 125:517–530

    Google Scholar 

  • Hegemann JH, Fleig UN (1993) The centromere of budding yeast. BioEssays 15:451–460

    Google Scholar 

  • Hegemann JH, Shero JH, Cottarel G, Philippsen P, Hieter P (1988) Mutational analysis of centromeric DNA from chromosome VI ofSaccharomyces cerevisiae. Mol Cell Biol 8: 2523–2535

    Google Scholar 

  • Hieter P, Pridmore D, Hegemann JH, Thomas M, Davis RW, Philippsen P (1985) Functional selection and analysis of yeast centromeric DNAs. Cell 42:913–921

    Google Scholar 

  • Huffaker TC, Thomas JH, Botstein D (1988) Diverse effects of β-tubulin mutations on microtubule formation and function. J Cell Biol 106:1997–2010

    Google Scholar 

  • Iida H, Yahara J (1984) Specific early-G1 blocks accompanied with stringent response inSaccharomyces cerevisiae lead to growth arrest in resting state similar to the G0 of higher cells. J Cell Biol 98:1185–1193

    Google Scholar 

  • Jiang W, Philippsen P (1989) Purification of a protein binding to the CDEI subregion ofSaccharomyces cerevisiae. Mol Cell Biol 9:5585–5593

    Google Scholar 

  • Jiang W, Lechner J, Carbon J (1993) Isolation and characterization of a gene (CBF2) specifying a protein component of the budding yeast kinetochore. J Cell Biol 121:513–519

    Google Scholar 

  • Johnston GC, Singer RA, McFarlane ES (1977) Growth and cell division during nitrogen starvation of the yeastSaccharomyces cerevisiae. J Bacteriol 132:723–730

    Google Scholar 

  • Kingsbury J, Koshland D (1993) Centromere-dependent binding of yeast minichromosomes to microtubules in vitro. Cell 66: 483–495

    Google Scholar 

  • Kingsbury J, Koshland D (1993) Centromere function on minichromosomes isolated from budding yeast. Mol Biol Cell 4: 859–870

    Google Scholar 

  • Lechner J (1994) A zinc finger protein, essential for chromosome segregation, constitutes a putative DNA binding subunit of theSaccharomyces cerevisiae kinetochore complex, Cbf3. EMBO J 13:5203–5211

    Google Scholar 

  • Lechner J, Carbon J (1991) A 240 kD multisubunit protein complex (CBF3) is a major component of the budding yeast centromere. Cell 64:717–727

    Google Scholar 

  • Marian B, Wintersberger U (1982) Modification of histones during the mitotic and meiotic cycle of yeast. FEBS Lett 139: 72–76

    Google Scholar 

  • McCarroll RM, Fangman WL (1988) Time of replication of yeast contromeres and telomeres. Cell 54:505–513

    Google Scholar 

  • Mellor J, Jiang W, Funk M, Rathjen J, Barnes CA, Hinz T, Hegemann JH, Philippsen P (1990) CPF1, a yeast protein which functions in centromeres and promoters. EMBO J 9: 4017–4026

    Google Scholar 

  • Niedenthal RK, Pick H, Sen-Gupta M, Wilmen A, Hegemann JH (1993) Cpfl protein induced bending of yeast centromeric DNA element I. Nucleic Acids Res 21:4726–4733

    Google Scholar 

  • Pinon R (1978) Folded chromosomes in non-cycling yeast cells. Chromosoma 67:263–274

    Google Scholar 

  • Saunders M, Fitzgerald-Hayes M, Bloom K (1988) Chromatin structure of altered yeast centromeres. Proc Natl Acad Sci USA 85:175–179

    Google Scholar 

  • Schmid A, Fascher K-D, Hörz W (1992) Nucleosome disruption at the yeastPHO5 promoter uponPHO5 induction occurs in the absence of DNA replication. Cell 71:853–864

    Google Scholar 

  • Schweizer B, Philippsen P (1991)CDC15, an essential cell cycle gene inSaccharomyces cerevisiae, encodes a protein kinase domain. Yeast 7:265–273

    Google Scholar 

  • Sethi N, Monteagudo MC, Koshland D, Hogan E, Burke DJ (1991) TheCDC20 gene product ofSaccharomyces cerevisiae, a β-transducin homolog, is required for a subset of microtubule-dependent cellular processes. Mol Cell Biol 11: 5592–5602

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1986) Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 163–167

    Google Scholar 

  • Spencer F, Hieter P (1992) Centromere DNA mutations induce a mitotic delay inS. cerevisiae. Proc Natl Acad Sci USA 89: 8908–8912

    Google Scholar 

  • Verhasselt P, Aert R, Voet M, Volckaert G (1994) Twelve open reading frames revealed on the 23.6 kb segment flanking the centromere on theSaccharomyces cerevisiae chromosome XIV right arm. YEAST 10:1355–1361

    Google Scholar 

  • Werner-Washburne M, Braun E, Johnston GC, Singer RA (1993) Stationary phase in the yeastSaccharomyces cerevisiae. Microbiol Rev 57:383–401

    Google Scholar 

  • Wilmen A (1994) Analysis of structural and functional components of theSaccharomyces cerevisiae centromeric complex. Ph.D thesis, University of Giessen, Giessen

  • Wood JS, Hartwell LH (1982) A dependent pathways of gene functions leading to chromosome segregation inSaccharomyces cerevisiae. J Cell Biol 94:718–726

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität, Frankfurter Strasse 107, D-35392, Giessen, Germany

    Andreas Wilmen & Johannes H. Hegemann

Authors
  1. Andreas Wilmen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Johannes H. Hegemann
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wilmen, A., Hegemann, J.H. The chromatin of theSaccharomyces cerevisiae centromere shows cell-type specific changes. Chromosoma 104, 489–503 (1996). https://doi.org/10.1007/BF00352113

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00352113

Keywords

Search

Navigation