scholarly journals Involvement of Sir2/4 in Silencing of DNA Breakage and Recombination on Mouse YACs during Yeast Meiosis

2005 ◽  
Vol 16 (3) ◽  
pp. 1449-1455 ◽  
Author(s):  
Yair Klieger ◽  
Ofer Yizhar ◽  
Drora Zenvirth ◽  
Neta Shtepel-Milman ◽  
Margriet Snoek ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Hot Spot ◽  
Yeast Cells ◽  
Class Iii ◽  
Dna Double Strand Breaks ◽  
Artificial Chromosomes ◽  
Mouse Major Histocompatibility Complex ◽  
Dna Backbone ◽  
Yeast Meiosis ◽  
Yeast Artificial Chromosomes

Yeast artificial chromosomes (YACs) that contain human DNA backbone undergo DNA double-strand breaks (DSBs) and recombination during yeast meiosis at rates similar to the yeast native chromosomes. Surprisingly, YACs containing DNA covering a recombination hot spot in the mouse major histocompatibility complex class III region do not show meiotic DSBs and undergo meiotic recombination at reduced levels. Moreover, segregation of these YACs during meiosis is seriously compromised. In meiotic yeast cells carrying the mutations sir2 or sir4, but not sir3, these YACs show DSBs, suggesting that a unique chromatin structure of the YACs, involving Sir2 and Sir4, protects the YACs from the meiotic recombination machinery. We speculate that the paucity of DSBs and recombination events on these YACs during yeast meiosis may reflect the refractory nature of the corresponding region in the mouse genome.

Recombination hot spot in the human beta-globin gene cluster: meiotic recombination of human DNA fragments in Saccharomyces cerevisiae

1985 ◽  
Vol 5 (8) ◽  
pp. 2029-2038
Author(s):  
D Treco ◽  
B Thomas ◽  
N Arnheim
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
High Frequency ◽  
Genetic Recombination ◽  
Hot Spot ◽  
Globin Gene ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Human Dna ◽  
Yeast Meiosis

We describe a novel system for the analysis of sequence-specific meiotic recombination in Saccharomyces cerevisiae. A comparison of three adjacent restriction fragments from the human beta-globin locus revealed that one of them, previously hypothesized to contain a relative hot spot for genetic recombination, engages in reciprocal exchange during yeast meiosis significantly more frequently than either of the other two fragments. Removal of the longest of four potential Z-DNA-forming regions from this fragment does not affect the high frequency of genetic recombination.

scholarly journals Recombination hot spot in the human beta-globin gene cluster: meiotic recombination of human DNA fragments in Saccharomyces cerevisiae.

1985 ◽  
Vol 5 (8) ◽  
pp. 2029-2038 ◽  
Author(s):  
D Treco ◽  
B Thomas ◽  
N Arnheim
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Meiotic Recombination ◽  
High Frequency ◽  
Genetic Recombination ◽  
Hot Spot ◽  
Globin Gene ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Human Dna ◽  
Yeast Meiosis

We describe a novel system for the analysis of sequence-specific meiotic recombination in Saccharomyces cerevisiae. A comparison of three adjacent restriction fragments from the human beta-globin locus revealed that one of them, previously hypothesized to contain a relative hot spot for genetic recombination, engages in reciprocal exchange during yeast meiosis significantly more frequently than either of the other two fragments. Removal of the longest of four potential Z-DNA-forming regions from this fragment does not affect the high frequency of genetic recombination.

scholarly journals Stable episomal maintenance of yeast artificial chromosomes in human cells.

1996 ◽  
Vol 16 (9) ◽  
pp. 5117-5126 ◽  
Author(s):  
K Simpson ◽  
A McGuigan ◽  
C Huxley
Keyword(s):  
Cell Lines ◽  
Viral Genome ◽  
Close Association ◽  
Human Cells ◽  
Yeast Cells ◽  
Control Of Gene Expression ◽  
Artificial Chromosomes ◽  
Barr Virus ◽  
Episomal Maintenance ◽  
Yeast Artificial Chromosomes

Plasmids carrying the Epstein-Barr virus origin of plasmid replication (oriP) have been shown to replicate autonomously in latently infected human cells (J. Yates, N. Warren, D. Reisman, and B. Sugden, Proc. Natl. Acad. Sci. USA 81:3806-3810, 1984). We demonstrate that addition of this domain is sufficient for stable episomal maintenance of yeast artificial chromosomes (YACs), up to at least 660 kb, in human cells expressing the viral protein EBNA-1. To better approximate the latent viral genome, YACs were circularized before addition of the oriP domain by homologous recombination in yeast cells. The resulting OriPYACs were maintained as extrachromosomal molecules over long periods in selection; a 90-kb OriPYAC was unrearranged in all cell lines analyzed, whereas the intact form of a 660-kb molecule was present in two of three cell lines. The molecules were also relatively stable in the absence of selection. This finding indicates that the oriP-EBNA-1 interaction is sufficient to stabilize episomal molecules of at least 660 kb and that such elements do not undergo rearrangements over time. Fluorescence in situ hybridization analysis demonstrated a close association of OriPYACs, some of which were visible as pairs, with host cell chromosomes, suggesting that the episomes replicate once per cell cycle and that stability is achieved by attachment to host chromosomes, as suggested for the viral genome. The wide availability of YAC libraries, the ease of manipulation of cloned sequences in yeast cells, and the episomal stability make OriPYACs ideal for studying gene function and control of gene expression.

scholarly journals Meiotic recombination on artificial chromosomes in yeast.

Genetics ◽  
1992 ◽  
Vol 131 (3) ◽  
pp. 541-550 ◽  
Author(s):  
L O Ross ◽  
D Treco ◽  
A Nicolas ◽  
J W Szostak ◽  
D Dawson
Keyword(s):  
Meiotic Recombination ◽  
Artificial Chromosome ◽  
Artificial Chromosomes ◽  
Crossover Behavior ◽  
Yeast Chromosome ◽  
Low Levels ◽  
Chromosome Iii ◽  
The Right ◽  
Yeast Artificial Chromosomes ◽  
Derivatives Of

Abstract We have examined the meiotic recombination characteristics of artificial chromosomes in Saccharomyces cerevisiae. Our experiments were carried out using minichromosome derivatives of yeast chromosome III and yeast artificial chromosomes composed primarily of bacteriophage lambda DNA. Tetrad analysis revealed that the artificial chromosomes exhibit very low levels of meiotic recombination. However, when a 12.5-kbp fragment from yeast chromosome VIII was inserted into the right arm of the artificial chromosome, recombination within that arm mimicked the recombination characteristics of the fragment in its natural context including the ability of crossovers to ensure meiotic disjunction. Both crossing over and gene conversion (within the ARG4 gene contained within the fragment) were measured in the experiments. Similarly, a 55-kbp region from chromosome III carried on a minichromosome showed crossover behavior indistinguishable from that seen when it is carried on chromosome III. We discuss the notion that, in yeast, meiotic recombination behavior is determined locally by small chromosomal regions that function free of the influence of the chromosome as a whole.

scholarly journals DNase I-hypersensitive sites and transcription factor-binding motifs within the mouse E beta meiotic recombination hot spot.

1991 ◽  
Vol 11 (4) ◽  
pp. 1813-1819 ◽  
Author(s):  
R Shenkar ◽  
M H Shen ◽  
N Arnheim
Keyword(s):  
Transcription Factors ◽  
Meiotic Recombination ◽  
Hot Spot ◽  
Dnase I ◽  
Binding Motifs ◽  
Dnase I Hypersensitive Sites ◽  
Mouse Major Histocompatibility Complex ◽  
Nuclear Extracts ◽  
Hypersensitive Sites ◽  
Beta Gene

The second intron of the E beta gene in the mouse major histocompatibility complex is the site of a meiotic recombination hot spot. We detected two DNase I-hypersensitive sites in this intron in meiotic cells isolated from mouse testes. One site appears to be constitutive and is found in other tissues regardless of whether or not they express the E beta gene. Near this hypersensitive site are potential binding motifs for H2TF1/KBF1, NF kappa B, and octamer transcription factors. Gel retardation studies with mouse lymphoma cell nuclear extracts confirmed that each of these motifs is capable of binding protein. The binding of transcription factors may contribute to the enhancement of recombination potential by altering chromatin structure and increasing the accessibility of the DNA to the recombination machinery.

scholarly journals Mps2 links Csm4 and Mps3 to form a telomere-associated LINC complex in budding yeast

2020 ◽  
Author(s):  
Jinbo Fan ◽  
Hui Jin ◽  
Bailey A. Koch ◽  
Hong-Guo Yu
Keyword(s):  
Nuclear Membrane ◽  
Meiotic Recombination ◽  
Budding Yeast ◽  
Transmembrane Proteins ◽  
Yeast Cells ◽  
Linc Complex ◽  
Domain Protein ◽  
Yeast Meiosis ◽  
Sun Domain ◽  
Variant Formation

AbstractThe linker of the nucleoskeleton and cytoskeleton (LINC) protein complex is composed of a pair of transmembrane proteins: the KASH-domain protein localized to the outer nuclear membrane and the SUN-domain protein to the inner nuclear membrane. In budding yeast, the sole SUN-domain protein, Mps3, is thought to pair with either Csm4 or Mps2, two KASH-like proteins, to form two separate LINC complexes. Here we show that Mps2 mediates the interaction between Csm4 and Mps3 to form a heterotrimeric telomere-associated LINC (t-LINC) in budding yeast meiosis. Mps2 binds to Csm4 and Mps3, and all three are localized to the telomere. Telomeric localization of Csm4 depends on both Mps2 and Mps3; in contrast, Mps2’s localization depends on Mps3 but not Csm4. Mps2-mediated t-LINC regulates telomere movement and meiotic recombination. By ectopically expressing CSM4 in vegetative yeast cells, we reconstitute the heterotrimeric t-LINC and demonstrate its ability to tether telomeres. Our findings therefore reveal the heterotrimeric composition of t-LINC in budding yeast and have implications for understanding LINC variant formation.

scholarly journals Mps2 links Csm4 and Mps3 to form a telomere-associated LINC complex in budding yeast

2020 ◽  
Vol 3 (12) ◽  
pp. e202000824
Author(s):  
Jinbo Fan ◽  
Hui Jin ◽  
Bailey A Koch ◽  
Hong-Guo Yu
Keyword(s):  
Nuclear Membrane ◽  
Meiotic Recombination ◽  
Budding Yeast ◽  
Transmembrane Proteins ◽  
Yeast Cells ◽  
The Sun ◽  
Linc Complex ◽  
Domain Protein ◽  
Yeast Meiosis ◽  
Sun Domain

The linker of the nucleoskeleton and cytoskeleton (LINC) complex is composed of two transmembrane proteins: the KASH domain protein localized to the outer nuclear membrane and the SUN domain protein to the inner nuclear membrane. In budding yeast, the sole SUN domain protein, Mps3, is thought to pair with either Csm4 or Mps2, two KASH-like proteins, to form two separate LINC complexes. Here, we show that Mps2 mediates the interaction between Csm4 and Mps3 to form a heterotrimeric telomere-associated LINC (t-LINC) complex in budding yeast meiosis. Mps2 binds to Csm4 and Mps3, and all three are localized to the telomere. Telomeric localization of Csm4 depends on both Mps2 and Mps3; in contrast, Mps2’s localization depends on Mps3 but not Csm4. Mps2-mediated t-LINC complex regulates telomere movement and meiotic recombination. By ectopically expressing CSM4 in vegetative yeast cells, we reconstitute the heterotrimeric t-LINC complex and demonstrate its ability to tether telomeres. Our findings therefore reveal the heterotrimeric composition of the t-LINC complex in budding yeast and have implications for understanding variant LINC complex formation.

DNase I-hypersensitive sites and transcription factor-binding motifs within the mouse E beta meiotic recombination hot spot

1991 ◽  
Vol 11 (4) ◽  
pp. 1813-1819
Author(s):  
R Shenkar ◽  
M H Shen ◽  
N Arnheim
Keyword(s):  
Transcription Factors ◽  
Meiotic Recombination ◽  
Hot Spot ◽  
Dnase I ◽  
Binding Motifs ◽  
Dnase I Hypersensitive Sites ◽  
Mouse Major Histocompatibility Complex ◽  
Nuclear Extracts ◽  
Hypersensitive Sites ◽  
Beta Gene

The second intron of the E beta gene in the mouse major histocompatibility complex is the site of a meiotic recombination hot spot. We detected two DNase I-hypersensitive sites in this intron in meiotic cells isolated from mouse testes. One site appears to be constitutive and is found in other tissues regardless of whether or not they express the E beta gene. Near this hypersensitive site are potential binding motifs for H2TF1/KBF1, NF kappa B, and octamer transcription factors. Gel retardation studies with mouse lymphoma cell nuclear extracts confirmed that each of these motifs is capable of binding protein. The binding of transcription factors may contribute to the enhancement of recombination potential by altering chromatin structure and increasing the accessibility of the DNA to the recombination machinery.

scholarly journals Meiotic recombination between yeast artificial chromosomes yields a single clone containing the entire BCL2 protooncogene.

1990 ◽  
Vol 87 (24) ◽  
pp. 9913-9917 ◽  
Author(s):  
G. A. Silverman ◽  
E. D. Green ◽  
R. L. Young ◽  
J. I. Jockel ◽  
P. H. Domer ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Artificial Chromosomes ◽  
Single Clone ◽  
Yeast Artificial Chromosomes
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