In eukaryotic cells, cohesin holds sister chromatids together until they separate into daughter cells during mitosis. We have used chromatin immunoprecipitation coupled with microarray analysis (ChIP chip) to produce a genome-wide description of cohesin binding to meiotic and mitotic chromosomes of Saccharomyces cerevisiae. A computer program, PeakFinder, enables flexible, automated identification and annotation of cohesin binding peaks in ChIP chip data. Cohesin sites are highly conserved in meiosis and mitosis, suggesting that chromosomes share a common underlying structure during different developmental programs. These sites occur with a semiperiodic spacing of 11 kb that correlates with AT content. The number of sites correlates with chromosome size; however, binding to neighboring sites does not appear to be cooperative. We observed a very strong correlation between cohesin sites and regions between convergent transcription units. The apparent incompatibility between transcription and cohesin binding exists in both meiosis and mitosis. Further experiments reveal that transcript elongation into a cohesin-binding site removes cohesin. A negative correlation between cohesin sites and meiotic recombination sites suggests meiotic exchange is sensitive to the chromosome structure provided by cohesin. The genome-wide view of mitotic and meiotic cohesin binding provides an important framework for the exploration of cohesins and cohesion in other genomes.
References
[1]
Baetz KK, Krogan NJ, et al. (2004) The ctf13–30/CTF13 genomic haploinsufficiency modifier screen identifies the yeast chromatin remodeling complex RSC, which is required for the establishment of sister chromatid cohesion. Mol Cell Biol 24: 1232–1244.
[2]
Bailey TL, Elkan C (1994) Fitting a mixture model by expectation maximization to discover motifs in biopolymers. Proc Int Conf Intell Syst Mol Biol 2: 28–36.
[3]
Bernard P, Maure JF, Partridge JF, Genier S, Javerzat JP, et al. (2001) Requirement of heterochromatin for cohesion at centromeres. Science 294: 2539–2542.
[4]
Blat Y, Kleckner N (1999) Cohesins bind to preferential sites along yeast chromosome III, with differential regulation along arms versus the centric region. Cell 98: 249–259.
[5]
Blat Y, Protacio RU, Hunter N, Kleckner N (2002) Physical and functional interactions among basic chromosome organizational features govern early steps of meiotic chiasma formation. Cell 111: 791–802.
[6]
Bohlander SK, Espinosa R, Le Beau MM, Rowley JD, Diaz MO (1992) A method for the rapid sequence-independent amplification of microdissected chromosomal material. Genomics 13: 1322–1324.
[7]
Buonomo SB, Clyne RK, Fuchs J, Loidl J, Uhlmann F, et al. (2000) Disjunction of homologous chromosomes in meiosis I depends on proteolytic cleavage of the meiotic cohesin Rec8 by separin. Cell 103: 387–398.
[8]
Bussemaker HJ, Li H, Siggia ED (2000) Building a dictionary for genomes: Identification of presumptive regulatory sites by statistical analysis. Proc Natl Acad Sci U S A 97: 10096–10100.
[9]
Chu S, DeRisi J, Eisen M, Mulholland J, Botstein D, et al. (1998) The transcriptional program of sporulation in budding yeast. Science 282: 699–705.
[10]
Cohen-Fix O, Koshland D (1997) The anaphase inhibitor of Pds1p is a target of the DNA damage checkpoint pathway. Proc Natl Acad Sci U S A 94: 14361–14366.
[11]
Gerton JL, DeRisi J, Shroff R, Lichten M, Brown PO, et al. (2000) Inaugural article: Global mapping of meiotic recombination hotspots and coldspots in the yeast . Proc Natl Acad Sci U S A 97: 11383–11390.
[12]
Green ED, Braden VV, Fulton RS, Lim R, Ueltzen MS, et al. (1995) A human chromosome 7 yeast artificial chromosome (YAC) resource: Construction, characterization, and screening. Genomics 25: 170–183.
[13]
Gruber S, Haering CH, Nasmyth K (2003) Chromosomal cohesin forms a ring. Cell 112: 765–777.
[14]
Guacci V, Koshland D, Strunnikov A (1997) A direct link between sister chromatid cohesion and chromosome condensation revealed through the analysis of MCD1 in . Cell 91: 47–57.
[15]
Hagstrom KA, Meyer BJ (2003) Condensin and cohesin: More than chromosome compactor and glue. Nat Rev Genet 4: 520–534.
[16]
Hakimi MA, Bochar DA, Schmiesing JA, Dong Y, Barak OG, et al. (2002) A chromatin remodelling complex that loads cohesin onto human chromosomes. Nature 418: 994–998.
[17]
Hill A, Bloom K (1987) Genetic manipulation of centromere function. Mol Cell Biol 7: 2397–2405.
[18]
Huang J, Hsu JM, et al. (2004) The RSC nucleosome-remodeling complex is required for cohesin's association with chromosome arms. Mol Cell 13: 739–750.
[19]
Hughes JD, Estep PW, Tavazoie S, Church GM (2000) Computational identification of cis-regulatory elements associated with groups of functionally related genes in . J Mol Biol 296: 1205–1214.
[20]
Iyer VR, Horak CE, Scafe CS, Botstein D, Snyder M, Brown PO (2001) Genomic binding sites of the yeast cell-cycle transcription factors SBF and MBF. Nature 409: 533–538.
[21]
Kitajima TS, Yokobayashi S, Yamamoto M, Watanabe Y (2003) Distinct cohesin complexes organize meiotic chromosome domains. Science 300: 1152–1155.
[22]
Klein F, Mahr P, Galova M, Buonomo SB, Michaelis C, et al. (1999) A central role for cohesins in sister chromatid cohesion, formation of axial elements, and recombination during yeast meiosis. Cell 98: 91–103.
[23]
Laloraya S, Guacci V, Koshland D (2000) Chromosomal addresses of the cohesin component Mcd1p. J Cell Biol 151: 1047–1056.
[24]
Lee CK, Shibata Y, Rao B, Strahl BD, Lieb JD (2004) Evidence for nucleosome depletion at active regulatory regions genome-wide. Nat Genet 36: In press.
[25]
Lieb JD, Liu X, Botstein D, Brown PO (2001) Promoter-specific binding of Rap1 revealed by genome-wide maps of protein-DNA association. Nat Genet 28: 327–334.
[26]
Liu XS, Brutlag DL, Liu JS (2002) An algorithm for finding protein-DNA binding sites with applications to chromatin-immunoprecipitation microarray experiments. Nat Biotechnol 20: 835–839.
[27]
Megee PC, Mistrot C, Guacci V, Koshland D (1999) The centromeric sister chromatid cohesion site directs Mcd1p binding to adjacent sequences. Mol Cell 4: 445–450.
[28]
Meluh PB, Koshland D (1997) Budding yeast centromere composition and assembly as revealed by in vivo cross-linking. Genes Dev 11: 3401–3412.
[29]
Milutinovich M, Koshland DE (2003) Molecular biology. SMC complexes—wrapped up in controversy. Science 300: 1101–1102.
[30]
Nagy PL, Cleary ML, Brown PO, Lieb JD (2003) Genomewide demarcation of RNA polymerase II transcription units revealed by physical fractionation of chromatin. Proc Natl Acad Sci U S A 100: 6364–6369.
[31]
Nonaka N, Kitajima T, Yokobayashi S, Xiao G, Yamamoto M, et al. (2002) Recruitment of cohesin to heterochromatic regions by Swi6/HP1 in fission yeast. Nat Cell Biol 4: 89–93.
[32]
Parker R, Herrick D, Peltz SW, Jacobson A (1991) Measurement of mRNA decay rates in . Methods Enzymol 194: 415–423.
[33]
Petes TD, Botstein D (1977) Simple Mendelian inheritance of the reiterated ribosomal DNA of yeast. Proc Natl Acad Sci U S A 74: 5091–5095.
[34]
Poloumienko A, Dershowitz A, De J, Newlon CS (2001) Completion of replication map of chromosome III. Mol Biol Cell 12: 3317–3327.
[35]
Reynolds AE, McCarroll RM, Newlon CS, Fangman WL (1989) Time of replication of ARS elements along yeast chromosome III. Mol Cell Biol 9: 4488–4494.
[36]
Rollins RA, Morcillo P, Dorsett D (1999) Nipped-B, a Drosophila homologue of chromosomal adherins, participates in activation by remote enhancers in the cut and Ultrabithorax genes. Genetics 152: 577–593.
San-Segundo PA, Roeder GS (2000) Role for the silencing protein Dot1 in meiotic checkpoint control. Mol Biol Cell 11: 3601–3615.
[39]
Snyder M, Sapolsky RJ, Davis RW (1988) Transcription interferes with elements important for chromosome maintenance in . Mol Cell Biol 8: 2184–2194.
[40]
Tanaka T, Cosma MP, Wirth K, Nasmyth K (1999) Identification of cohesin association sites at centromeres and along chromosome arms. Cell 98: 847–858.
[41]
Tanaka T, Fuchs J, Loidl J, Nasmyth K (2000) Cohesin ensures bipolar attachment of microtubules to sister centromeres and resists their precocious separation. Nat Cell Biol 2: 492–499.
[42]
Uhlmann F, Nasmyth K (1998) Cohesion between sister chromatids must be established during DNA replication. Curr Biol 8: 1095–1101.
[43]
Wade C, Shea KA, Jensen RV, McAlear MA (2001) EBP2 is a member of the yeast RRB regulon, a transcriptionally coregulated set of genes that are required for ribosome and rRNA biosynthesis. Mol Cell Biol 21: 8638–8650.
[44]
Watanabe Y, Nurse P (1999) Cohesin Rec8 is required for reductional chromosome segregation at meiosis. Nature 400: 461–464.
[45]
Weber SA, Gerton JL, Polancic JE, DeRisi JL, Koshland D, et al. (2004) The kinetochore is an enhancer of pericentric cohesion binding. PLoS Biol 2(9): e260. doi: 10.1371/journal.pbio.0020260.
[46]
Wu X, Wu C, Haber JE (1997) Rules of donor preference in Saccharomyces mating-type gene switching revealed by a competition assay involving two types of recombination. Genetics 147: 399–407.
[47]
Wyrick JJ, Aparicio JG, Chen T, Barnett JD, Jennings EG, et al. (2001) Genome-wide distribution of ORC and MCM proteins in S. cerevisiae high-resolution mapping of replication origins. Science 294: 2357–2360.
[48]
Yamamoto A, Guacci V, Koshland D (1996) Pds1p, an inhibitor of anaphase in budding yeast, plays a critical role in the APC and checkpoint pathway(s). J Cell Biol 133: 99–110.