Functional dissection of in vivo interchromosome association in Saccharomyces cerevisiae

Aragón-Alcaide, Luis; Strunnikov, Alexander

doi:10.1038/35041055

Cited by 44 publications

(28 citation statements)

References 42 publications

(7 reference statements)

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“…Coactivated genes can also form "transcription factories" or "multigene complexes" in higher eukaryotic species (33,34), so our findings here are likely to be one example of a more general phenomenon. This result can also explain why there have been conflicting observations on homologous pairing in yeast (12)(13)(14)(15): The experimental outcome likely depends on the inspected regions and their transcription status. The molecular nature of the interallelic interaction requires further elucidation.…”

Section: Discussionmentioning

confidence: 86%

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Interallelic interaction and gene regulation in budding yeast

Zhang

Bai

2016

Proc. Natl. Acad. Sci. U.S.A.

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In Drosophila, homologous chromosome pairing leads to "transvection," in which the enhancer of a gene can regulate the allelic transcription in trans. Interallelic interactions were also observed in vegetative diploid budding yeast, but their functional significance is unknown. Here, we show that a GAL1 reporter can interact with its homologous allele and affect its expression. By ectopically inserting two allelic reporters, one driven by wild-type GAL1 promoter (WT GAL1pr) and the other by a mutant promoter with delayed response to galactose induction, we found that the two reporters physically associate, and the WT GAL1pr triggers synchronized firing of the defective promoter and accelerates its activation without affecting its steady-state expression level. This interaction and the transregulatory effect disappear when the same reporters are located at nonallelic sites. We further demonstrated that the activator Gal4 is essential for the interallelic interaction, and the transregulation requires fully activated WT GAL1pr transcription. The mechanism of this phenomenon was further discussed. Taken together, our data revealed the existence of interallelic gene regulation in yeast, which serves as a starting point for understanding long-distance gene regulation in this genetically tractable system. homologous pairing | interallelic interaction | interallelic regulation | transvection | single-cell gene expression

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Section: Discussionmentioning

confidence: 86%

“…Homologous pairing was also observed in diploid budding yeast during vegetative growth, although the extent of the pairing is controversial among literature (12)(13)(14)(15). Unlike the Drosophila homologous chromosomes that stably associate with each other, the yeast chromosomes tend to interact more dynamic and sporadic at interstitial locations (12,14).…”

mentioning

confidence: 92%

Interallelic interaction and gene regulation in budding yeast

Zhang

Bai

2016

Proc. Natl. Acad. Sci. U.S.A.

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“…The main reason for the increase in allelic and ectopic association frequency of the lac operator (compared to the flanking sequences in wild-type conditions) is most likely the repetitive nature of the transgene construct. Sequence-specific but more or less locationindependent somatic association of multiple inserted arrays of tet operator and lac operator has been reported for budding yeast (Aragon-Alcaide and Strunnikov, 2000), although this was not confirmed by FISH or in the absence of fusion protein.…”

Section: Fish Analyses Of the Transgenic Line El702c With Flankingmentioning

confidence: 99%

“…The GFP-lac repressor protein is either transiently or stably expressed. The system was applied to various eukaryotes such as yeasts (Aragon-Alcaide and Strunnikov, 2000;Nabeshima et al, 1998;Straight et al, 1996), flies (Gunawardena and Rykowski, 2000;Vazquez et al, 2002), cultured mammalian cells Tsukamoto et al, 2000) and plants (Esch et al, 2003;Kato and Lam, 2001;Matzke et al, 2003). It revealed structural dynamics of chromosomes in interphase as well as mitotic cells by tracing the tagged loci in living cells and contributed to the understanding of chromosome functions (reviewed by Belmont et al, 1999;Gasser, 2002;Lam et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Tandem repetitive transgenes and fluorescent chromatin tags alter local interphase chromosome arrangement in Arabidopsis thaliana

Pečinka

Kato

Meister

et al. 2005

Journal of Cell Science

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Fluorescent protein chromatin tagging as achieved by the lac operator/lac repressor system is useful to trace distinct chromatin domains in living eukaryotic nuclei. To interpret the data correctly, it is important to recognize influences of the tagging system on nuclear architecture of the host cells. Within an Arabidopsis line that carries lac operator/lac repressor/GFP transgenes, the transgene loci frequently associate with each other and with heterochromatic chromocenters. Accumulation of tagged fusion protein further enhances the association frequency. Independent experiments with a transgenic plant carrying another multi-copy transgene also revealed, independent of its transcriptional state, unusually high frequencies of association with each other and with heterochromatin. From these results we conclude that the lac operator/lac repressor chromatin tagging system may alter the spatial chromatin organization in the host nuclei (in particular when more than one insertion locus is present) and also that loci of homologous transgenic repeats associate more often with each other and with endogenous heterochromatin than normal euchromatic regions.

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“…This implies the existence of a homology search mechanism as part of the recombination process. The integration of E. coli lacO and tetO sequences into chromosomes of yeast strains encoding lac repressor-GFP and tet repressor-GFP fusion proteins has recently provided visual evidence of long-distance interactions between like sequences (Aragon- Alcaide and Strunnikov, 2000). Such interactions may represent a constitutive homology search process.…”

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confidence: 99%

Nuclear oscillations and nuclear filament formation accompany single-strand annealing repair of a dicentric chromosome in Saccharomyces cerevisiae

Thrower

Yeh

Bloom

2003

Journal of Cell Science

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IntroductionThe study of DNA repair mechanisms has been aided by the development of experimental systems that generate DNA double-strand breaks (DSBs) at specific chromosomal locations and during defined periods of the cell cycle. One such system is the budding yeast conditional dicentric chromosome (Hill and Bloom, 1989). This inducible dicentric chromosome was constructed by inserting a second copy of centromere 3 (CEN3) into chromosome III, approximately 45 kb upstream of the endogenous centromere 3 (Fig. 1A). The inserted centromere is under control of the GAL1 promoter, allowing for the reversible inactivation of this centromere when cells are grown in the presence of galactose. Studies utilizing dicentric chromosomes are unique in that DSBs are generated by mitotic pulling forces when centromeres on the same sister chromatid are aligned to opposite spindle poles during mitosis. Previous work showed that efficient healing of a broken dicentric chromosome in Saccharomyces cerevisiae requires RAD52, indicating that repair occurs primarily through homologous recombination . In cells lacking RAD52, infrequent dicentric chromosome repair events occur through non-homologous end joining (Kramer et al., 1994).Resolution of a conditional dicentric chromosome is accompanied by the deletion of one centromere and the intervening DNA. These events were initially thought to result from gene conversion (GC) accompanied by crossover (Jager and Philippsen, 1989). However, other studies of direct repeat recombination have shown that deletions commonly occur without generating the circular product predicted to form through a crossover event (Fishman-Lobell et al., 1992). Moreover, many deletion events are dependent on RAD1 and RAD10 (Klein, 1988;Schiestl and Prakash, 1990;Ivanov and Haber, 1995). The Rad1 and Rad10 proteins form a structurespecific endonuclease that generates 5′ single-stranded ends (Siede et al., 1993). These genes function in a recombination pathway known as single-strand annealing (SSA) (Sugawara and Haber, 1992;Sung et al., 1993;Ivanov and Haber, 1995). The recognition of SSA as a major pathway for direct repeat recombination suggests that this mechanism plays a role in the repair of a dicentric chromosome.Previous studies have shown that recombination frequency is primarily governed by the degree of sequence homology rather than the location of the recombining sequences within the genome (Haber and Leung, 1996). This implies the existence of a homology search mechanism as part of the recombination process. The integration of E. coli lacO and tetO sequences into chromosomes of yeast strains encoding lac repressor-GFP and tet repressor-GFP fusion proteins has recently provided visual evidence of long-distance interactions between like sequences (Aragon- Alcaide and Strunnikov, 2000). Such interactions may represent a constitutive homology search process. The mechanisms that bring homologous sequences into close association are unknown. Efficient meiotic recombination in the fission yeast Schizosaccharomyce...

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Functional dissection of in vivo interchromosome association in Saccharomyces cerevisiae

Cited by 44 publications

References 42 publications

Interallelic interaction and gene regulation in budding yeast

Interallelic interaction and gene regulation in budding yeast

Tandem repetitive transgenes and fluorescent chromatin tags alter local interphase chromosome arrangement in Arabidopsis thaliana

Nuclear oscillations and nuclear filament formation accompany single-strand annealing repair of a dicentric chromosome in Saccharomyces cerevisiae

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