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Recombination protein Tid1p controls resolution of cohesin-dependent linkages in meiosis in Saccharomyces cerevisiae.

Kateneva AV, Konovchenko AA, Guacci V, Dresser ME - J. Cell Biol. (2005)

Bottom Line: Genetic results indicate that the primary defect in these cells is a failure to resolve Mcd1p-mediated connections.Tid1p interacts with recombination enzymes Dmc1p and Rad51p and has an established role in recombination repair.We propose that Tid1p remodels Mcd1p-mediated cohesion early in meiotic prophase to facilitate interhomologue recombination and the subsequent segregation of homologous chromosomes.

View Article: PubMed Central - PubMed

Affiliation: Program in Molecular, Cell, and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104.

ABSTRACT
Sister chromatid cohesion and interhomologue recombination are coordinated to promote the segregation of homologous chromosomes instead of sister chromatids at the first meiotic division. During meiotic prophase in Saccharomyces cerevisiae, the meiosis-specific cohesin Rec8p localizes along chromosome axes and mediates most of the cohesion. The mitotic cohesin Mcd1p/Scc1p localizes to discrete spots along chromosome arms, and its function is not clear. In cells lacking Tid1p, which is a member of the SWI2/SNF2 family of helicase-like proteins that are involved in chromatin remodeling, Mcd1p and Rec8p persist abnormally through both meiotic divisions, and chromosome segregation fails in the majority of cells. Genetic results indicate that the primary defect in these cells is a failure to resolve Mcd1p-mediated connections. Tid1p interacts with recombination enzymes Dmc1p and Rad51p and has an established role in recombination repair. We propose that Tid1p remodels Mcd1p-mediated cohesion early in meiotic prophase to facilitate interhomologue recombination and the subsequent segregation of homologous chromosomes.

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Suppression of tid1Δ sister chromatid separation defect by mcd1-1 in a spo11Δ spo13Δ background. (A) Percentage of cells with two separated chromatin masses (binucleates) after a shift to nonpermissive (33°C) temperature at 2-h intervals. Cells were allowed to sporulate for at least 24 h after the shift to nonpermissive temperature before scoring. Fig. S3 (A and B; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1) and (C) show the kinetics of the short spindle, dumbbell, and binucleate stages at 21°C during this experiment. (B and C) Percentage of binucleates (B) and dumbbells (C) after a shift from 21 to 33°C at 4 h. (D) Percentage of cells with short spindles after a shift from 21 to 33°C at 4 h. Progression of cells through the division is monitored by the behavior of the spindle (TUB1-GFP) and DAPI-stained chromatin. At least 200 cells were scored for each time point.
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fig5: Suppression of tid1Δ sister chromatid separation defect by mcd1-1 in a spo11Δ spo13Δ background. (A) Percentage of cells with two separated chromatin masses (binucleates) after a shift to nonpermissive (33°C) temperature at 2-h intervals. Cells were allowed to sporulate for at least 24 h after the shift to nonpermissive temperature before scoring. Fig. S3 (A and B; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1) and (C) show the kinetics of the short spindle, dumbbell, and binucleate stages at 21°C during this experiment. (B and C) Percentage of binucleates (B) and dumbbells (C) after a shift from 21 to 33°C at 4 h. (D) Percentage of cells with short spindles after a shift from 21 to 33°C at 4 h. Progression of cells through the division is monitored by the behavior of the spindle (TUB1-GFP) and DAPI-stained chromatin. At least 200 cells were scored for each time point.

Mentions: In mitotic cells, the maintenance of cohesion between sister chromatids requires Mcd1p, which is a homologue of Rec8p (Guacci et al., 1997; Michaelis et al., 1997). Although Mcd1p has been reported not to have a major role in the maintenance of cohesion during meiosis in budding yeast (Klein et al., 1999), it does play a role in meiotic sister chromatid cohesion in fission yeast (Yokobayashi et al., 2003). Therefore, we tested whether Mcd1p is required for persistent sister chromatid connections in tid1Δ by using a heat-sensitive allele of MCD1 (mcd1-1), which was previously shown to be defective in cohesion maintenance at nonpermissive temperature in mitotic cells (Guacci et al., 1997). Progression of cells through the division was monitored by using TUB1-GFP and DAPI-stained chromatin. Sporulating cells were shifted from permissive for mcd1-1 temperature (21°C) to nonpermissive temperature (33°C) every 2 h, and binucleates were scored after at least 24 h of sporulation. Early shift to 33°C completely rescued the tid1Δ anaphase block (compare spo11Δ spo13Δ mcd1-1 with tid1Δ vs. without tid1Δ; Fig. 5 A). However, this suppression disappears by metaphase (compare spo11Δ spo13Δ mcd1-1 tid1Δ in Fig. 5 A with spo11Δ spo13Δ mcd1-1 in Fig. S3 C, in which cells with short spindles represent metaphase cells; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1). To more precisely determine the effect of Mcd1p inactivation on the tid1Δ anaphase block, the percentage of binucleates, dumbbells, and cells with a short spindle was compared in cells sporulating at 21°C and in cells shifted to 33°C at 4 h (Fig. 5, B–D, and Fig. S3, A–C), which is a time when mcd1-1 completely suppresses tid1Δ. Consistently, identical levels of binucleates are reached by spo11Δ spo13Δ mcd1-1 and spo11Δ spo13Δ mcd1-1 tid1Δ after a shift to 33°C at 4 h of sporulation. No accumulation of cells with a short spindle or of dumbbells was observed in spo11Δ spo13Δ mcd1-1 tid1Δ cells compared with spo11Δ spo13Δ mcd1-1. At 21°C, the phenotype of tid1Δ is unchanged by mcd1-1. These data suggest a direct involvement of Mcd1p in generating persistent connections in tid1Δ.


Recombination protein Tid1p controls resolution of cohesin-dependent linkages in meiosis in Saccharomyces cerevisiae.

Kateneva AV, Konovchenko AA, Guacci V, Dresser ME - J. Cell Biol. (2005)

Suppression of tid1Δ sister chromatid separation defect by mcd1-1 in a spo11Δ spo13Δ background. (A) Percentage of cells with two separated chromatin masses (binucleates) after a shift to nonpermissive (33°C) temperature at 2-h intervals. Cells were allowed to sporulate for at least 24 h after the shift to nonpermissive temperature before scoring. Fig. S3 (A and B; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1) and (C) show the kinetics of the short spindle, dumbbell, and binucleate stages at 21°C during this experiment. (B and C) Percentage of binucleates (B) and dumbbells (C) after a shift from 21 to 33°C at 4 h. (D) Percentage of cells with short spindles after a shift from 21 to 33°C at 4 h. Progression of cells through the division is monitored by the behavior of the spindle (TUB1-GFP) and DAPI-stained chromatin. At least 200 cells were scored for each time point.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2171180&req=5

fig5: Suppression of tid1Δ sister chromatid separation defect by mcd1-1 in a spo11Δ spo13Δ background. (A) Percentage of cells with two separated chromatin masses (binucleates) after a shift to nonpermissive (33°C) temperature at 2-h intervals. Cells were allowed to sporulate for at least 24 h after the shift to nonpermissive temperature before scoring. Fig. S3 (A and B; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1) and (C) show the kinetics of the short spindle, dumbbell, and binucleate stages at 21°C during this experiment. (B and C) Percentage of binucleates (B) and dumbbells (C) after a shift from 21 to 33°C at 4 h. (D) Percentage of cells with short spindles after a shift from 21 to 33°C at 4 h. Progression of cells through the division is monitored by the behavior of the spindle (TUB1-GFP) and DAPI-stained chromatin. At least 200 cells were scored for each time point.
Mentions: In mitotic cells, the maintenance of cohesion between sister chromatids requires Mcd1p, which is a homologue of Rec8p (Guacci et al., 1997; Michaelis et al., 1997). Although Mcd1p has been reported not to have a major role in the maintenance of cohesion during meiosis in budding yeast (Klein et al., 1999), it does play a role in meiotic sister chromatid cohesion in fission yeast (Yokobayashi et al., 2003). Therefore, we tested whether Mcd1p is required for persistent sister chromatid connections in tid1Δ by using a heat-sensitive allele of MCD1 (mcd1-1), which was previously shown to be defective in cohesion maintenance at nonpermissive temperature in mitotic cells (Guacci et al., 1997). Progression of cells through the division was monitored by using TUB1-GFP and DAPI-stained chromatin. Sporulating cells were shifted from permissive for mcd1-1 temperature (21°C) to nonpermissive temperature (33°C) every 2 h, and binucleates were scored after at least 24 h of sporulation. Early shift to 33°C completely rescued the tid1Δ anaphase block (compare spo11Δ spo13Δ mcd1-1 with tid1Δ vs. without tid1Δ; Fig. 5 A). However, this suppression disappears by metaphase (compare spo11Δ spo13Δ mcd1-1 tid1Δ in Fig. 5 A with spo11Δ spo13Δ mcd1-1 in Fig. S3 C, in which cells with short spindles represent metaphase cells; available at http://www.jcb.org/cgi/content/full/jcb.200505020/DC1). To more precisely determine the effect of Mcd1p inactivation on the tid1Δ anaphase block, the percentage of binucleates, dumbbells, and cells with a short spindle was compared in cells sporulating at 21°C and in cells shifted to 33°C at 4 h (Fig. 5, B–D, and Fig. S3, A–C), which is a time when mcd1-1 completely suppresses tid1Δ. Consistently, identical levels of binucleates are reached by spo11Δ spo13Δ mcd1-1 and spo11Δ spo13Δ mcd1-1 tid1Δ after a shift to 33°C at 4 h of sporulation. No accumulation of cells with a short spindle or of dumbbells was observed in spo11Δ spo13Δ mcd1-1 tid1Δ cells compared with spo11Δ spo13Δ mcd1-1. At 21°C, the phenotype of tid1Δ is unchanged by mcd1-1. These data suggest a direct involvement of Mcd1p in generating persistent connections in tid1Δ.

Bottom Line: Genetic results indicate that the primary defect in these cells is a failure to resolve Mcd1p-mediated connections.Tid1p interacts with recombination enzymes Dmc1p and Rad51p and has an established role in recombination repair.We propose that Tid1p remodels Mcd1p-mediated cohesion early in meiotic prophase to facilitate interhomologue recombination and the subsequent segregation of homologous chromosomes.

View Article: PubMed Central - PubMed

Affiliation: Program in Molecular, Cell, and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104.

ABSTRACT
Sister chromatid cohesion and interhomologue recombination are coordinated to promote the segregation of homologous chromosomes instead of sister chromatids at the first meiotic division. During meiotic prophase in Saccharomyces cerevisiae, the meiosis-specific cohesin Rec8p localizes along chromosome axes and mediates most of the cohesion. The mitotic cohesin Mcd1p/Scc1p localizes to discrete spots along chromosome arms, and its function is not clear. In cells lacking Tid1p, which is a member of the SWI2/SNF2 family of helicase-like proteins that are involved in chromatin remodeling, Mcd1p and Rec8p persist abnormally through both meiotic divisions, and chromosome segregation fails in the majority of cells. Genetic results indicate that the primary defect in these cells is a failure to resolve Mcd1p-mediated connections. Tid1p interacts with recombination enzymes Dmc1p and Rad51p and has an established role in recombination repair. We propose that Tid1p remodels Mcd1p-mediated cohesion early in meiotic prophase to facilitate interhomologue recombination and the subsequent segregation of homologous chromosomes.

Show MeSH
Related in: MedlinePlus