Lack of chromosome territoriality in yeast: promiscuous rejoining of broken chromosome ends.

Various studies suggest that eukarytoic chromosomes may occupy distinct territories within the nucleus and that chromosomes are tethered to a nuclear matrix. These constraints might limit interchromosomal interactions. We have used a molecular genetic test to investigate whether the chromosomes of Saccharomyces cerevisiae exhibit such territoriality. A chromosomal double-strand break (DSB) can be efficiently repaired by recombination between flanking homologous repeated sequences. We have constructed a strain in which DSBs are delivered simultaneously to both chromosome III and chromosome V by induction of the HO endonuclease. The arrangement of partially duplicated HIS4 and URA3 sequences around each HO recognition site allows the repair of the two DSBs in two alternative ways: (i) the creation of two intrachromosomal deletions or (ii) the formation of a pair of reciprocal translocations. We show that reciprocal translocations are formed approximately as often as the pair of intrachromosomal deletions. Similar results were obtained when one of the target regions was moved from chromosome V to any of three different locations on chromosome XI. These results argue that the broken ends of mitotic chromosomes are free to search the entire genome for appropriate partners; thus, mitotic chromosomes are not functionally confined to isolated domains of the nucleus, at least when chromosomes are broken.

[1]  J. Haber,et al.  Rad52-independent mitotic gene conversion in Saccharomyces cerevisiae frequently results in chromosomal loss. , 1985, Genetics.

[2]  S. Gasser,et al.  Chromosomal ARS and CEN elements bind specifically to the yeast nuclear scaffold , 1988, Cell.

[3]  V. Guacci,et al.  Chromosome condensation and sister chromatid pairing in budding yeast , 1994, The Journal of cell biology.

[4]  I. Herskowitz,et al.  Directionality and regulation of cassette substitution in yeast. , 1984, Cold Spring Harbor symposia on quantitative biology.

[5]  B. Dujon,et al.  Site-specific recombination determined by I-SceI, a mitochondrial group I intron-encoded endonuclease expressed in the yeast nucleus. , 1992, Genetics.

[6]  R. Rothstein,et al.  The genetic control of direct-repeat recombination in Saccharomyces: the effect of rad52 and rad1 on mitotic recombination at GAL10, a transcriptionally regulated gene. , 1989, Genetics.

[7]  J. Haber,et al.  Intermediates of recombination during mating type switching in Saccharomyces cerevisiae. , 1990, The EMBO journal.

[8]  D. Carroll,et al.  DNA recombination and repair in oocytes, eggs, and extracts. , 1991, Methods in cell biology.

[9]  N. Sternberg,et al.  Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process , 1984, Molecular and cellular biology.

[10]  D. Agard,et al.  The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis , 1993, The Journal of cell biology.

[11]  S. Gasser,et al.  DNA loops: structural and functional properties of scaffold‐attached regions , 1992, Molecular microbiology.

[12]  H. Klein Different types of recombination events are controlled by the RAD1 and RAD52 genes of Saccharomyces cerevisiae. , 1988, Genetics.

[13]  J. Haber,et al.  Two alternative pathways of double-strand break repair that are kinetically separable and independently modulated , 1992, Molecular and cellular biology.

[14]  R H Borts,et al.  Meiotic gene conversion and crossing over between dispersed homologous sequences occurs frequently in Saccharomyces cerevisiae. , 1987, Genetics.

[15]  J. Nickoloff,et al.  Double-strand breaks stimulate alternative mechanisms of recombination repair. , 1989, Journal of molecular biology.

[16]  E. Gilson,et al.  SIR3 and SIR4 proteins are required for the positioning and integrity of yeast telomeres , 1993, Cell.

[17]  S. Jinks-Robertson,et al.  Substrate length requirements for efficient mitotic recombination in Saccharomyces cerevisiae. , 1993, Molecular and cellular biology.

[18]  J. Haber,et al.  Characterization of double-strand break-induced recombination: homology requirements and single-stranded DNA formation , 1992, Molecular and cellular biology.

[19]  U. K. Laemmli,et al.  Scaffold-associated regions: cis-acting determinants of chromatin structural loops and functional domains. , 1992, Current opinion in genetics & development.

[20]  S. Gasser Replication origins, factors and attachment sites. , 1991, Current opinion in cell biology.

[21]  N. Sternberg,et al.  Repair of double-stranded DNA breaks by homologous DNA fragments during transfer of DNA into mouse L cells , 1990, Molecular and cellular biology.

[22]  L. Manuelidis A view of interphase chromosomes , 1990, Science.

[23]  J. Haber In vivo biochemistry: Physical monitoring of recombination induced by site‐specific endonucleases , 1995, BioEssays : news and reviews in molecular, cellular and developmental biology.