scholarly journals Sister chromatid repair maintains genomic integrity during meiosis in Caenorhabditis elegans

2020 ◽  
Author(s):  
Erik Toraason ◽  
Cordell Clark ◽  
Anna Horacek ◽  
Marissa L. Glover ◽  
Alina Salagean ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Sister Chromatid ◽  
Meiotic Prophase ◽  
Genome Integrity ◽  
Early Prophase ◽  
Dna Breaks ◽  
Late Prophase ◽  
Exogenous Dna ◽  
Prophase I ◽  
Meiotic Prophase I

SummaryDuring meiosis, the maintenance of genome integrity is critical for generating viable haploid gametes [1]. In meiotic prophase I, double-strand DNA breaks (DSBs) are induced and a subset of these DSBs are repaired as interhomolog crossovers to ensure proper chromosome segregation. DSBs in excess of the permitted number of crossovers must be repaired by other pathways to ensure genome integrity [2]. To determine if the sister chromatid is engaged for meiotic DSB repair during oogenesis, we developed an assay to detect sister chromatid repair events at a defined DSB site during Caenorhabditis elegans meiosis. Using this assay, we directly demonstrate that the sister chromatid is available as a meiotic repair template for both crossover and noncrossover recombination, with noncrossovers being the predominant recombination outcome. We additionally find that the sister chromatid is the exclusive recombination partner for DSBs during late meiotic prophase I. Analysis of noncrossover conversion tract sequences reveals that DSBs are processed similarly throughout prophase I and recombination intermediates remain central around the DSB site. Further, we demonstrate that the SMC-5/6 complex is required for long conversion tracts in early prophase I and intersister crossovers during late meiotic prophase I; whereas, the XPF-1 nuclease is required only in late prophase to promote sister chromatid repair. In response to exogenous DNA damage at different stages of meiosis, we find that mutants for SMC-5/6 and XPF-1 have differential effects on progeny viability. Overall, we propose that SMC-5/6 both processes recombination intermediates and promotes sister chromatid repair within meiotic prophase I, while XPF-1 is required as an intersister resolvase only in late prophase I.

scholarly journals Meiosis-specific prophase-like pathway controls cleavage-independent release of cohesin by Wapl phosphorylation

2018 ◽  
Author(s):  
Kiran Challa ◽  
V Ghanim Fajish ◽  
Miki Shinohara ◽  
Franz Klein ◽  
Susan M. Gasser ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Sister Chromatid ◽  
Meiotic Prophase ◽  
Budding Yeast ◽  
Cell Cycle Regulators ◽  
Late Prophase ◽  
Homologous Chromosomes ◽  
Prophase I ◽  
Meiotic Prophase I ◽  
Meiotic Chromosomes

AbstractSister chromatid cohesion on chromosome arms is essential for the segregation of homologous chromosomes during meiosis I while it is dispensable for sister chromatid separation during mitosis. It was assumed that, unlike the situation in mitosis, chromosome arms retain cohesion prior to onset of anaphase-I. Paradoxically, reduced immunostaining signals of meiosis-specific cohesin, including the kleisin Rec8, from the chromosomes were observed during late prophase-I of budding yeast. This decrease is seen in the absence of Rec8 cleavage and depends on condensin-mediated recruitment of Polo-like kinase (PLK/Cdc5). In this study, we confirmed that this release indeed accompanies the dissociation of acetylated Smc3 as well as Rec8 from meiotic chromosomes during late prophase-I. This release requires, in addition to PLK, the cohesin regulator, Wapl (Rad61/Wpl1 in yeast), and Dbf4-dependent Cdc7 kinase (DDK). Meiosis-specific phosphorylation of Rad61/Wpl1 and Rec8 by PLK and DDK collaboratively promote this release. This process is similar to the vertebrate “prophase” pathway for cohesin release during G2 phase and pro-metaphase. In yeast, meiotic cohesin release coincides with PLK-dependent compaction of chromosomes in late meiotic prophase-I. We suggest that yeast uses this highly regulated cleavage-independent pathway to remove cohesin during late prophase-I to facilitate morphogenesis of condensed metaphase-I chromosomes.Author SummaryIn meiosis the life and health of future generations is decided upon. Any failure in chromosome segregation has a detrimental impact. Therefore, it is currently believed that the physical connections between homologous chromosomes are maintained by meiotic cohesin with exceptional stability. Indeed, it was shown that cohesive cohesin does not show an appreciable turnover during long periods in oocyte development. In this context, it was long assumed but not properly investigated, that the prophase pathway for cohesin release would be specific to mitotic cells and will be safely suppressed during meiosis so as not to endanger the valuable chromosome connections. However, a previous study on budding yeast meiosis suggests the presence of cleavage-independent pathway of cohesin release during late prophase-I. In the work presented here we confirmed that the prophase pathway is not suppressed during meiosis, at least in budding yeast and showed that this cleavage-independent release is regulated by meiosis-specific phosphorylation of two cohesin subunits, Rec8 and Rad61(Wapl) by two cell-cycle regulators, PLK and DDK. Our results suggest that late meiotic prophase-I actively controls cohesin dynamics on meiotic chromosomes for chromosome segregation.

scholarly journals The CSN/COP9 Signalosome Regulates Synaptonemal Complex Assembly during Meiotic Prophase I of Caenorhabditis elegans

PLoS Genetics ◽  
2014 ◽  
Vol 10 (11) ◽  
pp. e1004757 ◽  
Author(s):  
Heather Brockway ◽  
Nathan Balukoff ◽  
Martha Dean ◽  
Benjamin Alleva ◽  
Sarit Smolikove
Keyword(s):  
Caenorhabditis Elegans ◽  
Synaptonemal Complex ◽  
Meiotic Prophase ◽  
Cop9 Signalosome ◽  
Complex Assembly ◽  
Prophase I ◽  
Meiotic Prophase I

scholarly journals akirin is required for diakinesis bivalent structure and synaptonemal complex disassembly at meiotic prophase I

2013 ◽  
Vol 24 (7) ◽  
pp. 1053-1067 ◽  
Author(s):  
Amy M. Clemons ◽  
Heather M. Brockway ◽  
Yizhi Yin ◽  
Bhavatharini Kasinathan ◽  
Yaron S. Butterfield ◽  
...  
Keyword(s):  
Synaptonemal Complex ◽  
Meiotic Prophase ◽  
Chromosome Condensation ◽  
Late Prophase ◽  
Homologous Chromosomes ◽  
Prophase I ◽  
Meiotic Prophase I ◽  
Evolutionarily Conserved ◽  
Key Events

During meiosis, evolutionarily conserved mechanisms regulate chromosome remodeling, leading to the formation of a tight bivalent structure. This bivalent, a linked pair of homologous chromosomes, is essential for proper chromosome segregation in meiosis. The formation of a tight bivalent involves chromosome condensation and restructuring around the crossover. The synaptonemal complex (SC), which mediates homologous chromosome association before crossover formation, disassembles concurrently with increased condensation during bivalent remodeling. Both chromosome condensation and SC disassembly are likely critical steps in acquiring functional bivalent structure. The mechanisms controlling SC disassembly, however, remain unclear. Here we identify akir-1 as a gene involved in key events of meiotic prophase I in Caenorhabditis elegans. AKIR-1 is a protein conserved among metazoans that lacks any previously known function in meiosis. We show that akir-1 mutants exhibit severe meiotic defects in late prophase I, including improper disassembly of the SC and aberrant chromosome condensation, independently of the condensin complexes. These late-prophase defects then lead to aberrant reconfiguring of the bivalent. The meiotic divisions are delayed in akir-1 mutants and are accompanied by lagging chromosomes. Our analysis therefore provides evidence for an important role of proper SC disassembly in configuring a functional bivalent structure.

scholarly journals Exposure to the anthelmintic dinitroaniline oryzalin causes changes in meiotic prophase morphology and loss of synaptonemal complexes in the nematode Caenorhabditis elegans

2021 ◽  
Vol 2 ◽  
Author(s):  
Paul Goldstein
Keyword(s):  
Caenorhabditis Elegans ◽  
Nuclear Matrix ◽  
Meiotic Prophase ◽  
Parasitic Nematodes ◽  
Nematode Caenorhabditis Elegans ◽  
Synaptonemal Complexes ◽  
Prophase I ◽  
Meiotic Prophase I

Abstract The anthelmintic dinitroaniline oryzalin interferes with the formation of microtubules and inhibits meiosis and mitosis in nematodes. Exposure to oryzalin resulted in deterioration in morphology of the oocytes and loss of synaptonemal complexes at meiotic prophase I. The nuclear matrix and envelope were poorly formed, and the central rachis was diminished. These results provide the basis for the loss of fecundity after treatment with the oryzalin resulting in control of parasitic nematodes.

scholarly journals Sex chromosome pairing mediated by euchromatic homology in Drosophila male meiosis

2019 ◽  
Author(s):  
Christopher A. Hylton ◽  
Katie Hansen ◽  
Andrew Bourgeois ◽  
John E. Tomkiel
Keyword(s):  
Sex Chromosomes ◽  
Sex Chromosome ◽  
Intergenic Spacer ◽  
Male Meiosis ◽  
Early Prophase ◽  
Dna Breaks ◽  
Late Prophase ◽  
Prophase I ◽  
Y Chromosomes ◽  
Meiosis I

ABSTRACTTo maintain proper ploidy, haploid sex cells must undergo two subsequent meiotic divisions. During meiosis I, homologs pair and remain conjoined until segregation at anaphase. Drosophila melanogaster spermatocytes are unique in that the canonical events of meiosis I including synaptonemal complex (SC) formation, double-strand DNA breaks, and chiasmata are absent. Sex chromosomes pair at intergenic spacer sequences within the heterochromatic rDNA while euchromatin is required to pair and segregate autosomal homologies, suggesting that pairing may be limited to specific sequences. However, previous work generated from genetic segregation assays or observations of late prophase I/prometaphase I chromosome associations fail to differentiate pairing from conjunction. Here, we separately examined the capability of X euchromatin to pair and conjoin using an rDNA-deficient X and a series of Dp(1;Y) chromosomes. Genetic assays showed that duplicated X euchromatin can substitute for endogenous rDNA pairing sites. Segregation was not proportional to homology length, and pairing could be mapped to nonoverlapping sequences within a single Dp(1;Y). Using fluorescent in situ hybridization (FISH) to early prophase I spermatocytes, we showed that pairing occurred with high fidelity at all homologies tested. Pairing was unaffected by the presence of X rDNA, nor could it be explained by rDNA magnification. By comparing genetic and cytological data, we determined that centromere proximal pairings were best at segregation. Segregation was dependent on the conjunction protein Stromalin in Meiosis while the autosomal-specific Teflon was dispensable. Overall, our results suggest that pairing may occur at all homologies, but there may be sequence or positional requirements for conjunction.ARTICLE SUMMARYDrosophila males have evolved a unique system of chromosome segregation in meiosis that lacks recombination. Chromosomes pair at selected sequences suggesting that early steps of meiosis may also differ in this organism. Using Y chromosomes carrying portions of X material, we show that pairing between sex chromosomes can be mediated by sequences other than the previously identified rDNA pairing sites. We propose that pairing may simply be homology-based and may not differ from canonical meiosis observed in females. The main difference in males may be that conjunctive mechanisms that join homologs in the absence of crossovers.

scholarly journals A mutation in the endonuclease domain of mouse MLH3 reveals novel roles for MutLγ during crossover formation in meiotic prophase I

2019 ◽  
Author(s):  
Melissa Toledo ◽  
Xianfei Sun ◽  
Miguel A. Brieño-Enríquez ◽  
Vandana Raghavan ◽  
Stephen Gray ◽  
...  
Keyword(s):  
Meiotic Prophase ◽  
Early Prophase ◽  
Crossing Over ◽  
Dsb Repair ◽  
Normal Loading ◽  
Strand Breaks ◽  
Prophase I ◽  
Meiotic Prophase I ◽  
Endonuclease Domain ◽  
Blm Helicase

ABSTRACTDuring meiotic prophase I, double-strand breaks (DSBs) initiate homologous recombination leading to non-crossovers (NCOs) and crossovers (COs). In mouse, 10% of DSBs are designated to become COs, primarily through a pathway dependent on the MLH1-MLH3 heterodimer (MutLγ). Mlh3 contains an endonuclease domain that is critical for resolving COs in yeast. We generated a mouse (Mlh3DN/DN) harboring a mutation within this conserved domain that is predicted to generate a protein that is catalytically inert. Mlh3DN/DN males, like fully null Mlh3-/- males, have no spermatozoa and are infertile, yet spermatocytes have normal DSBs and undergo normal synapsis events in early prophase I. Unlike Mlh3-/- males, mutation of the endonuclease domain within MLH3 permits normal loading and frequency of MutLγ in pachynema. However, key DSB repair factors (RAD51) and mediators of CO pathway choice (BLM helicase) persist into pachynema in Mlh3DN/DN males, indicating a temporal delay in repair events and revealing a mechanism by which alternative DSB repair pathways may be selected. While Mlh3DN/DN spermatocytes retain only 22% of wildtype chiasmata counts, this frequency is greater than observed in Mlh3-/- males (10%), suggesting that the allele may permit partial endonuclease activity, or that other pathways can generate COs from these MutLγ-defined repair intermediates in Mlh3DN/DN males. Double mutant mice homozygous for the Mlh3DN/DN and Mus81-/- mutations show losses in chiasmata that approach levels observed in Mlh3-/- males, indicating that the MUS81-EME1-regulated crossover pathway accounts for some of the increased residual chiasmata observed in Mlh3DN/DN spermatocytes. Our data demonstrate that mouse spermatocytes bearing the MLH1-MLH3DN/DN complex display the proper loading of factors essential for CO resolution (MutSγ, CDK2, HEI10, MutLγ). Despite these functions, mice bearing the Mlh3DN/DN allele show defects in the repair of meiotic recombination intermediates and a loss of most chiasmata.SUMMARYThe MLH1-MLH3 complex is essential for crossing over in mammalian meiosis. We generated a mutation in mouse MLH3 that alters its conserved endonuclease domain and show that it disrupts crossing over in a manner distinct from the full null Mlh3 mouse, but also results in male infertility.

scholarly journals Sex Chromosome Pairing Mediated by Euchromatic Homology in Drosophila Male Meiosis

Genetics ◽  
2020 ◽  
Vol 214 (3) ◽  
pp. 605-616 ◽  
Author(s):  
Christopher A. Hylton ◽  
Katie Hansen ◽  
Andrew Bourgeois ◽  
John E. Tomkiel Dean
Keyword(s):  
Sex Chromosome ◽  
Intergenic Spacer ◽  
Male Meiosis ◽  
Early Prophase ◽  
Dna Breaks ◽  
Late Prophase ◽  
Prophase I ◽  
Double Strand Dna Breaks ◽  
Chromosome Associations ◽  
Meiosis I

Diploid germline cells must undergo two consecutive meiotic divisions before differentiating as haploid sex cells. During meiosis I, homologs pair and remain conjoined until segregation at anaphase. Drosophila melanogaster spermatocytes are unique in that the canonical events of meiosis I including synaptonemal complex formation, double-strand DNA breaks, and chiasmata are absent. Sex chromosomes pair at intergenic spacer sequences within the ribosomal DNA (rDNA). Autosomes pair at numerous euchromatic homologies, but not at heterochromatin, suggesting that pairing may be limited to specific sequences. However, previous work generated from genetic segregation assays or observations of late prophase I/prometaphase I chromosome associations fail to differentiate pairing from maintenance of pairing (conjunction). Here, we separately examined the capability of X euchromatin to pair and conjoin using an rDNA-deficient X and a series of Dp(1;Y) chromosomes. Genetic assays showed that duplicated X euchromatin can substitute for endogenous rDNA pairing sites. Segregation was not proportional to homology length, and pairing could be mapped to nonoverlapping sequences within a single Dp(1;Y). Using fluorescence in situ hybridization to early prophase I spermatocytes, we showed that pairing occurred with high fidelity at all homologies tested. Pairing was unaffected by the presence of X rDNA, nor could it be explained by rDNA magnification. By comparing genetic and cytological data, we determined that centromere proximal pairings were best at segregation. Segregation was dependent on the conjunction protein Stromalin in Meiosis, while the autosomal-specific Teflon was dispensable. Overall, our results suggest that pairing may occur at all homologies, but there may be sequence or positional requirements for conjunction.

scholarly journals Efficient Repair of DNA Damage Induced by Heavy Ion Particles in Meiotic Prophase I Nuclei of Caenorhabditis elegans

2003 ◽  
Vol 44 (3) ◽  
pp. 271-276 ◽  
Author(s):  
TAKAKO TAKANAMI ◽  
YONGZHAO ZHANG ◽  
HIDETOSHI AOKI ◽  
TOMOKO ABE ◽  
SHIGEO YOSHIDA ◽  
...  
Keyword(s):  
Dna Damage ◽  
Caenorhabditis Elegans ◽  
Meiotic Prophase ◽  
Heavy Ion ◽  
Prophase I ◽  
Meiotic Prophase I

scholarly journals Automated and customizable quantitative image analysis of whole Caenorhabditis elegans germlines

Genetics ◽  
2021 ◽  
Author(s):  
Erik Toraason ◽  
Victoria L Adler ◽  
Nicole A Kurhanewicz ◽  
Acadia DiNardo ◽  
Adam M Saunders ◽  
...  
Keyword(s):  
Image Analysis ◽  
Caenorhabditis Elegans ◽  
Germ Cell ◽  
Meiotic Prophase ◽  
Quantitative Image Analysis ◽  
Dna Breaks ◽  
C Elegans ◽  
Quantitative Image ◽  
Cytological Features ◽  
Single Nucleus

Abstract Arranged in a spatial-temporal gradient for germ cell development, the adult germline of Caenorhabditis elegans is an excellent system for understanding the generation, differentiation, function, and maintenance of germ cells. Imaging whole C. elegans germlines along the distal-proximal axis enables powerful cytological analyses of germ cell nuclei as they progress from the pre-meiotic tip through all the stages of meiotic prophase I. To enable high-content image analysis of whole C. elegans gonads, we developed a custom algorithm and pipelines to function with image processing software that enables: (1) quantification of cytological features at single nucleus resolution from immunofluorescence images; and (2) assessment of these individual nuclei based on their position within the germline. We show the capability of our quantitative image analysis approach by analyzing multiple cytological features of meiotic nuclei in whole C. elegans germlines. First, we quantify double-strand DNA breaks (DSBs) per nucleus by analyzing DNA-associated foci of the recombinase RAD-51 at single-nucleus resolution in the context of whole germline progression. Second, we quantify the DSBs that are licensed for crossover repair by analyzing foci of MSH-5 and COSA-1 when they associate with the synaptonemal complex during meiotic prophase progression. Finally, we quantify P-granule composition across the whole germline by analyzing the colocalization of PGL-1 and ZNFX-1 foci. Our image analysis pipeline is an adaptable and useful method for researchers spanning multiple fields using the C. elegans germline as a model system.

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