Loss of sister kinetochore co-orientation and peri-centromeric cohesin protection after meiosis I depends on cleavage of centromeric REC8

Construction of strains to identify novel factors for regulation of centromeric cohesion protection (CCP) and sister kinetochore mono-orientation (SKM)

BMC Molecular and Cell Biology ◽

10.1186/s12860-019-0231-2 ◽

2019 ◽

Vol 20 (1) ◽

Cited By ~ 1

Author(s):

Akhilendra Pratap Bharati ◽

Santanu Kumar Ghosh

Keyword(s):

Budding Yeast ◽

Cdna Libraries ◽

The Novel ◽

Unique Type ◽

Sister Kinetochore ◽

Single Protein ◽

Dividing Cells ◽

Specific Factors ◽

Meiosis I ◽

Segregation Of Chromosomes

Abstract Background Meiosis-I is a unique type of chromosome segregation where each chromosome aligns and segregates from its homolog. The mechanism of meiosis I homolog separation in different eukaryotes depends on their centromere and kinetochore architecture which in turn relies mainly on two processes, first on a specialized four protein complex known as monopolin and second, the centromeric cohesion protection (CCP). However, in mammals the complex has not been identified. Furthermore, in budding yeast, there could be additional factors in this process which includes some meiosis specific and some non meiosis specific factors. Result We constructed two strains. In the first strain we expressed Mam1 and Cdc5 which leads to sister kinetochore monoorientation (SKM) and in the second case we expressed Rec8 and Spo13 which enhanced CCP even in mitosis. The expression of these proteins in mitotically dividing cells caused co-orientation of the chromosomes, which lead to the cell death followed by miss-segregation of chromosomes. Then we utilized these strains to screen the cDNA libraries from yeast and mammals to identify the novel factors which participate in CCP and SKM. Finally, SGY4119 strain expressing Spo13 and Rec8 was transformed with pRS316 gal cDNA library and transformants were screened for lethality on galactose. We screened ~ 105 transformants colonies. Out of these ~ 3000 colonies were able to survive on galactose plate which was narrow down to 6 on the basis of desired phenotype. Conclusion So far, meiosis specific kinetochore proteins have been identified only in two yeasts. Recently, in mammals a meiosis specific kinetochore protein (MEIKIN) has been identified with similar function. Till now a single protein in mammals and four proteins monopolin complex in budding yeast has been identified to coorient the centromere. Many more novel factors have to be identified yet. That is why we wished to device genetic screen using a functional genomics approach. Since the list of proteins already identified in yeast is not exhaustive as the circumstantial evidence suggests, we wish to use the same yeast strains to identify additional novel yeast proteins that may be involved in the execution of meiosis.

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Aurora controls sister kinetochore mono-orientation and homolog bi-orientation in meiosis-I

The EMBO Journal ◽

10.1038/sj.emboj.7601880 ◽

2007 ◽

Vol 26 (21) ◽

pp. 4475-4486 ◽

Cited By ~ 61

Author(s):

Silke Hauf ◽

Ashapurno Biswas ◽

Maria Langegger ◽

Shigehiro A Kawashima ◽

Tatsuya Tsukahara ◽

...

Keyword(s):

Sister Kinetochore ◽

Meiosis I

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Shugoshin Promotes Sister Kinetochore Biorientation in Saccharomyces cerevisiae

Molecular Biology of the Cell ◽

10.1091/mbc.e07-06-0584 ◽

2008 ◽

Vol 19 (3) ◽

pp. 1199-1209 ◽

Cited By ~ 36

Author(s):

Brendan M. Kiburz ◽

Angelika Amon ◽

Adele L. Marston

Keyword(s):

Saccharomyces Cerevisiae ◽

Chromosome Segregation ◽

Minor Role ◽

Meiotic Cell ◽

Homologous Chromosomes ◽

Cell Divisions ◽

Sister Chromatids ◽

Sister Kinetochore ◽

A Minor ◽

Meiosis I

Chromosome segregation must be executed accurately during both mitotic and meiotic cell divisions. Sgo1 plays a key role in ensuring faithful chromosome segregation in at least two ways. During meiosis this protein regulates the removal of cohesins, the proteins that hold sister chromatids together, from chromosomes. During mitosis, Sgo1 is required for sensing the absence of tension caused by sister kinetochores not being attached to microtubules emanating from opposite poles. Here we describe a differential requirement for Sgo1 in the segregation of homologous chromosomes and sister chromatids. Sgo1 plays only a minor role in segregating homologous chromosomes at meiosis I. In contrast, Sgo1 is important to bias sister kinetochores toward biorientation. We suggest that Sgo1 acts at sister kinetochores to promote their biorientation.

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Spo13 prevents premature cohesin cleavage during meiosis

Wellcome Open Research ◽

10.12688/wellcomeopenres.15066.2 ◽

2019 ◽

Vol 4 ◽

pp. 29 ◽

Cited By ~ 6

Author(s):

Stefan Galander ◽

Rachael E. Barton ◽

David A. Kelly ◽

Adèle L. Marston

Keyword(s):

Regulatory Subunit ◽

Homologous Chromosomes ◽

Sister Chromatids ◽

Functional Homolog ◽

Sister Kinetochore ◽

Phosphatase 2A ◽

Cohesin Subunit ◽

Yeast Meiosis ◽

Mitosis And Meiosis ◽

Meiosis I

Background: Meiosis produces gametes through two successive nuclear divisions, meiosis I and meiosis II. In contrast to mitosis and meiosis II, where sister chromatids are segregated, during meiosis I, homologous chromosomes are segregated. This requires the monopolar attachment of sister kinetochores and the loss of cohesion from chromosome arms, but not centromeres, during meiosis I. The establishment of both sister kinetochore mono-orientation and cohesion protection rely on the budding yeast meiosis I-specific Spo13 protein, the functional homolog of fission yeast Moa1 and mouse MEIKIN. Methods: Here we investigate the effects of loss of SPO13 on cohesion during meiosis I using a live-cell imaging approach. Results: Unlike wild type, cells lacking SPO13 fail to maintain the meiosis-specific cohesin subunit, Rec8, at centromeres and segregate sister chromatids to opposite poles during anaphase I. We show that the cohesin-destabilizing factor, Wpl1, is not primarily responsible for the loss of cohesion during meiosis I. Instead, premature loss of centromeric cohesin during anaphase I in spo13Δ cells relies on separase-dependent cohesin cleavage. Further, cohesin loss in spo13Δ anaphase I cells is blocked by forcibly tethering the regulatory subunit of protein phosphatase 2A, Rts1, to Rec8. Conclusions: Our findings indicate that separase-dependent cleavage of phosphorylated Rec8 causes premature cohesin loss in spo13Δ cells.

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An interplay between Shugoshin and Spo13 for centromeric cohesin protection and sister kinetochore mono-orientation during meiosis I in Saccharomyces cerevisiae

Current Genetics ◽

10.1007/s00294-018-0832-x ◽

2018 ◽

Vol 64 (5) ◽

pp. 1141-1152 ◽

Cited By ~ 3

Author(s):

Gunjan Mehta ◽

Guhan Kaliyaperumal Anbalagan ◽

Akhilendra Pratap Bharati ◽

Purna Gadre ◽

Santanu Kumar Ghosh

Keyword(s):

Saccharomyces Cerevisiae ◽

Sister Kinetochore ◽

Meiosis I

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Complex regulation of sister kinetochore orientation in meiosis-I

Journal of Biosciences ◽

10.1007/s12038-010-0053-z ◽

2010 ◽

Vol 35 (3) ◽

pp. 485-495 ◽

Cited By ~ 7

Author(s):

Amit Bardhan

Keyword(s):

Sister Kinetochore ◽

Complex Regulation ◽

Meiosis I

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Spo13 prevents premature cohesin cleavage during meiosis

10.1101/488312 ◽

2018 ◽

Author(s):

Stefan Galander ◽

Rachael E Barton ◽

David A Kelly ◽

Adele L Marston

Keyword(s):

Regulatory Subunit ◽

Homologous Chromosomes ◽

Sister Chromatids ◽

Functional Homolog ◽

Sister Kinetochore ◽

Phosphatase 2A ◽

Cohesin Subunit ◽

Yeast Meiosis ◽

Mitosis And Meiosis ◽

Meiosis I

Meiosis produces gametes through two successive nuclear divisions, meiosis I and meiosis II. In contrast to mitosis and meiosis II, where sister chromatids are segregated, during meiosis I, homologous chromosomes are segregated. This requires the monopolar attachment of sister kinetochores and the loss of cohesion from chromosome arms, but not centromeres, during meiosis I. The establishment of both sister kinetochore mono-orientation and cohesion protection rely on the budding yeast meiosis I-specific Spo13 protein, the functional homolog of fission yeast Moa1 and mouse MEIKIN. Here we investigate the effects of loss of SPO13 on cohesion during meiosis I. Unlike wild type, cells lacking SPO13 fail to maintain the meiosis-specific cohesin subunit, Rec8, at centromeres and segregate sister chromatids to opposite poles during anaphase I. We show that the cohesin-destabilizing factor, Wpl1, is not primarily responsible for the loss of cohesion during meiosis I. Instead, premature loss of centromeric cohesin during anaphase I in spo13Δ cells relies on separase-dependent cohesin cleavage. Further, cohesin loss in spo13Δ anaphase I cells is blocked by forcibly tethering the regulatory subunit of protein phosphatase 2A, Rts1, to Rec8. Our findings indicate that separase-dependent cleavage of phosphorylated Rec8 causes premature cohesin loss in spo13Δ cells.

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Loss of Sister Kinetochore Co-orientation and Peri-centromeric Cohesin Protection after Meiosis I Depends on Cleavage of Centromeric REC8

10.1101/2020.02.06.935171 ◽

2020 ◽

Cited By ~ 2

Author(s):

Sugako Ogushi ◽

Ahmed Rattani ◽

Jonathan Godwin ◽

Jean Metson ◽

Lothar Schermelleh ◽

...

Keyword(s):

Catalytic Activity ◽

Sister Chromatid ◽

Spindle Pole ◽

Sister Chromatids ◽

Sister Kinetochore ◽

Meiosis I

SummaryProtection of peri-centromeric REC8 cohesin from separase and sister kinetochore attachment to microtubules emanating from the same spindle pole (co-orientation) ensure that sister chromatids remain associated after meiosis I. Both features are lost during meiosis II, when sister kinetochores bi-orient and lose peri-centromeric REC8 protection, resulting in sister chromatid disjunction and the production of haploid gametes. By transferring spindle-chromosome complexes (SCCs) between meiosis I and II cells, we have discovered that both sister kinetochore co-orientation and peri-centromeric cohesin protection depend on the SCC and not the cytoplasm. Moreover, the catalytic activity of separase at meiosis I is necessary not only for converting kinetochores from a co-to a bi-oriented state but also for deprotection of peri-centromeric cohesin and that cleavage of REC8 may be the key event. Crucially, we show that selective cleavage of REC8 in the vicinity of kinetochores is sufficient to destroy co-orientation in univalent chromosomes, albeit not in bivalents where resolution of chiasmata through cleavage of Rec8 along chromosome arms may also be required.

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Direct evaluation of cohesin-mediated sister kinetochore associations at meiosis I in fission yeast

Journal of Cell Science ◽

10.1242/jcs.259102 ◽

2021 ◽

Author(s):

Masashi Nambu ◽

Atsuki Kishikawa ◽

Takatomi Yamada ◽

Kento Ichikawa ◽

Yunosuke Kira ◽

...

Keyword(s):

Fission Yeast ◽

Chromosome Segregation ◽

Mitotic Chromosomes ◽

Direct Evaluation ◽

Homologous Chromosomes ◽

Sister Chromatids ◽

Pheromone Signaling ◽

Sister Kinetochore ◽

Novel Method ◽

Meiosis I

Kinetochores drive chromosome segregation by mediating chromosome interactions with the spindle. In higher eukaryotes, sister kinetochores are separately positioned on opposite sides of sister centromeres during mitosis, but associate with each other during meiosis I. Kinetochore association facilitates the attachment of sister chromatids to the same pole, enabling the segregation of homologous chromosomes toward opposite poles. In the fission yeast, Schizosaccharomyces pombe, Rec8-containing meiotic cohesin is suggested to establish kinetochore associations by mediating cohesion of the centromere cores. However, cohesin-mediated kinetochore associations on intact chromosomes have never been demonstrated directly. Here, we describe a novel method for the direct evaluation of kinetochore associations on intact chromosomes in live S. pombe cells, and demonstrate that sister kinetochores and the centromere cores are positioned separately on mitotic chromosomes but associate with each other on meiosis I chromosomes. Furthermore, we demonstrate that kinetochore association depends on meiotic cohesin and the cohesin regulators, Moa1 and Mrc1, and requires mating-pheromone signaling for its establishment. These results confirm cohesin-mediated kinetochore association and its regulatory mechanisms, along with the usefulness of the developed method for its analysis.

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