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Yeast meiosis

Bo Zhang, Anna M Butler, Qian Shi, Siyuan Xing, Paul K Herman
P-bodies are liquid droplet-like compartments that lack a limiting membrane and are present in many eukaryotic cells. These structures contain specific sets of proteins and mRNAs at concentrations higher than that in the surrounding environment. Although highly conserved, the normal physiological roles of these ribonucleoprotein (RNP) granules remain poorly defined. Here, we report that P-bodies are required for the efficient completion of meiosis in the budding yeast, Saccharomyces cerevisiae P-bodies were found to be present during all phases of the meiotic program and to provide protection for the Hrr25/CK1 protein kinase, a key regulator of this developmental process...
June 18, 2018: Molecular and Cellular Biology
Sophie Atkinson, Samuel Marguerat, Danny Bitton, Francois Bachand, Maria Rodriguez-Lopez, Charalampos Rallis, Jean-Francois Lemay, Cristina Cotobal, Michal Malecki, Pawel Smialowski, Juan Mata, Philipp Korber, Jurg Bahler
Long non-coding RNAs (lncRNAs), which are longer than 200 nucleotides but often unstable, contribute a substantial and diverse portion to pervasive non-coding transcriptomes. Most lncRNAs are poorly annotated and understood, although several play important roles in gene regulation and diseases. Here we systematically uncover and analyse lncRNAs in Schizosaccharomyces pombe. Based on RNA-seq data from twelve RNA-processing mutants and nine physiological conditions, we identify 5775 novel lncRNAs, nearly 4-times the previously annotated lncRNAs...
June 18, 2018: RNA
Jingxun Chen, David McSwiggen, Elçin Ünal
Single molecule fluorescence in situ hybridization (smFISH) is a powerful technique to study gene expression in single cells due to its ability to detect and count individual RNA molecules. Complementary to deep sequencing-based methods, smFISH provides information about the cell-to-cell variation in transcript abundance and the subcellular localization of a given RNA. Recently, we have used smFISH to study the expression of the gene NDC80 during meiosis in budding yeast, in which two transcript isoforms exist and the short transcript isoform has its entire sequence shared with the long isoform...
May 25, 2018: Journal of Visualized Experiments: JoVE
Ye Hong, Maria Velkova, Nicola Silva, Marlène Jagut, Viktor Scheidt, Karim Labib, Verena Jantsch, Anton Gartner
Homologous recombination is essential for crossover (CO) formation and accurate chromosome segregation during meiosis. It is of considerable importance to work out how recombination intermediates are processed, leading to CO and non-crossover (NCO) outcome. Genetic analysis in budding yeast and Caenorhabditis elegans indicates that the processing of meiotic recombination intermediates involves a combination of nucleases and DNA repair enzymes. We previously reported that in C. elegans meiotic joint molecule resolution is mediated by two redundant pathways, conferred by the SLX-1 and MUS-81 nucleases, and by the HIM-6 Bloom helicase in conjunction with the XPF-1 endonuclease, respectively...
June 7, 2018: PLoS Genetics
Meenakshi Agarwal, Hui Jin, Melainia McClain, Jinbo Fan, Bailey A Koch, Sue L Jaspersen, Hong-Guo Yu
The budding yeast centrosome, often called the spindle pole body (SPB), nucleates microtubules for chromosome segregation during cell division. An appendage, called the half bridge, attaches to one side of the SPB and regulates SPB duplication and separation. Like DNA, the SPB is duplicated only once per cell cycle. During meiosis, however, after one round of DNA replication, two rounds of SPB duplication and separation are coupled with homolog segregation in meiosis I and sister-chromatid segregation in meiosis II...
May 30, 2018: Molecular Biology of the Cell
Touko Niimi, Taro Nakamura
The spindle pole body (SPB) plays a central role in spore plasma membrane formation in addition to its recognized role in microtubule organization. During meiosis, a biomembrane called the forespore membrane (FSM) is newly formed at the SPB. Although several SPB proteins essential for the initiation of FSM formation (meiotic SPB components) have been identified, the molecular mechanism is still unknown. Here, we report the isolation and functional characterization of Dms1 as a component of the SPB. We show that FSM formation does not initiate in dms1Δ cells...
2018: PloS One
Rokas Grigaitis, Aitor Susperregui, Philipp Wild, Joao Matos
The formation of stable interactions between chromosomes of maternal and paternal origin-homologs-is required for their segregation during meiosis. To achieve this, cells take advantage of the recombination machinery, which promotes formation of reciprocal interhomolog exchanges, called crossovers, from the repair of self-inflicted DNA breaks. Important genetic studies led to the identification of key enzymes that control meiotic recombination. However, characterization of their biochemical properties when purified from meiotic cultures has been difficult to achieve...
2018: Methods in Cell Biology
Michel F Guiraldelli, Anna Felberg, Luciana P Almeida, Aniruddha Parikh, Rodrigo O de Castro, Roberto J Pezza
Chromosome segregation errors during meiosis result in the formation of aneuploid gametes and are the leading cause of pregnancy loss and birth defects in humans. Proper chromosome segregation requires pairwise associations of maternal and paternal homologous chromosomes. Chiasmata, which are the cytological manifestations of crossovers (COs), provide a physical link that holds the homologs together as a pair, facilitating their orientation on the spindle at meiosis I. Although CO-promoting activities ensure a balanced number and position of COs, their identity and mechanism of action in mammals remain understudied...
May 2018: PLoS Genetics
Kayla Carpenter, Rachel Brietta Bell, Julius Yunus, Angelika Amon, Luke Edwin Berchowitz
Amyloids are fibrous protein assemblies that are often described as irreversible and intrinsically pathogenic. However, yeast cells employ amyloid-like assemblies of the RNA-binding protein Rim4 to control translation during meiosis. Here, we show that multi-site phosphorylation of Rim4 is critical for its regulated disassembly and degradation and that failure to clear Rim4 assemblies interferes with meiotic progression. Furthermore, we identify the protein kinase Ime2 to bring about Rim4 clearance via phosphorylation of Rim4's intrinsically disordered region...
May 7, 2018: Developmental Cell
Steven Boeynaems, Aaron D Gitler
Protein aggregation can be beneficial, with important biological functions, but must be somehow controlled. In this issue of Developmental Cell, Carpenter et al. (2018) uncover how a solid-like supermolecular protein assembly that regulates yeast meiosis is disassembled through phosphorylation of a disordered prion-like domain to control the timing of meiotic progression.
May 7, 2018: Developmental Cell
Ignacio Flor-Parra, Ana Belén Iglesias-Romero, Silvia Salas-Pino, Rafael Lucena, Juan Jimenez, Rafael R Daga
In metazoans, the nuclear envelope (NE) breakdown (NEBD) occurs during "open" mitosis and meiosis. In the fission yeast Schizosaccharomyces pombe, the mitosis and the first meiotic division (MI) are "closed," during which the NE is maintained. Intriguingly, during the second meiotic division (MII), the NE is also maintained, but nuclear and cytoplasmic molecules are mixed similarly to open mitosis, a phenomenon of unknown biological significance called "virtual" NEBD (vNEBD). Here, we show that importin-α-dependent nucleocytoplasmic transport regulates spindle disassembly late in anaphase B at MI, as previously reported for mitosis...
April 24, 2018: Cell Reports
Joseph M Varberg, Sue L Jaspersen
In fission yeast, the nuclear envelope (NE) remains intact during mitosis and meiosis I but is compromised during meiosis II. In this issue of Cell Reports, Flor-Parra et al. (2018) demonstrate that this NE alteration regulates meiosis II spindle disassembly and the ploidy of meiotic products.
April 24, 2018: Cell Reports
Weber Beringui Feitosa, Patricia L Morris
SUMOylation, a process of posttranslational modification of proteins by the Small Ubiquitin related Modifier (SUMO) family of proteins, is known to be involved in yeast and mammalian somatic cell-cycle regulation. However, the identities of the SUMO-modified oocyte targets are largely unknown and the functional role(s) for SUMOylation during mammalian oocyte maturation remains unclear. Based upon studies in non-germline cells, protein kinase B/AKT is a potential SUMOylation target in the mouse oocyte, where it plays an essential role in cell-cycle resumption and progression during maturation...
April 18, 2018: American Journal of Physiology. Cell Physiology
Simon David Brown, Olga Dorota Jarosinska, Alexander Lorenz
Hop1 is a component of the meiosis-specific chromosome axis and belongs to the evolutionarily conserved family of HORMA domain proteins. Hop1 and its orthologs in higher eukaryotes are a major factor in promoting double-strand DNA break formation and inter-homolog recombination. In budding yeast and mammals, they are also involved in a meiotic checkpoint kinase cascade monitoring the completion of double-strand DNA break repair. We used the fission yeast, Schizosaccharomyces pombe, which lacks a canonical synaptonemal complex to test whether Hop1 has a role beyond supporting the generation of double-strand DNA breaks and facilitating inter-homolog recombination events...
March 17, 2018: Current Genetics
Hardeep Kaur, Jasvinder S Ahuja, Michael Lichten
Proteins with potential roles in meiotic recombination often have essential or important functions during the mitotic cell cycle. In addition, proteins may have different functions at different times during meiosis. In such cases, it can be challenging to precisely determine protein function during meiosis using null or hypomorphic mutants. One example is the Sgs1-Top3-Rmi1 helicase-decatenase complex, which is required for normal vegetative growth and genome stability. In such cases, conditional loss-of-function mutants can be useful...
2018: Methods in Enzymology
Eleni P Mimitou, Scott Keeney
During meiosis, the specialized cell division giving rise to gametes, numerous DNA double-strand breaks (DSBs) are introduced at multiple places throughout the genome by the topoisomerase-like protein Spo11. Homologous recombination, a highly conserved DSB repair pathway, is employed for their repair and ensures the formation of chiasmata and the proper segregation of homologous chromosomes. In the initial steps of recombination, end resection takes place, wherein Spo11 is endonucleolytically released and the 5'-terminal strands of each DSB are exonucleolytically processed, exposing the ssDNA necessary to identify a homologous repair template...
2018: Methods in Enzymology
Shannon Owens, Shangming Tang, Neil Hunter
Homologous recombination is fundamental to sexual reproduction, facilitating accurate segregation of homologous chromosomes at the first division of meiosis, and creating novel allele combinations that fuel evolution. Following initiation of meiotic recombination by programmed DNA double-strand breaks (DSBs), homologous pairing and DNA strand exchange form joint molecule (JM) intermediates that are ultimately resolved into crossover and noncrossover repair products. Physical monitoring of the DNA steps of meiotic recombination in Saccharomyces cerevisiae (budding yeast) cultures undergoing synchronous meiosis has provided seminal insights into the molecular basis of meiotic recombination and affords a powerful tool for dissecting the molecular roles of recombination factors...
2018: Methods in Enzymology
Ze Cheng, George Maxwell Otto, Emily Nicole Powers, Abdurrahman Keskin, Philipp Mertins, Steven Alfred Carr, Marko Jovanovic, Gloria Ann Brar
To better understand the gene regulatory mechanisms that program developmental processes, we carried out simultaneous genome-wide measurements of mRNA, translation, and protein through meiotic differentiation in budding yeast. Surprisingly, we observed that the levels of several hundred mRNAs are anti-correlated with their corresponding protein products. We show that rather than arising from canonical forms of gene regulatory control, the regulation of at least 380 such cases, or over 8% of all measured genes, involves temporally regulated switching between production of a canonical, translatable transcript and a 5' extended isoform that is not efficiently translated into protein...
February 22, 2018: Cell
Fabien Moretto, N Ezgi Wood, Gavin Kelly, Andreas Doncic, Folkert J van Werven
Transcription of long noncoding RNAs (lncRNAs) regulates local gene expression in eukaryotes. Many examples of how a single lncRNA controls the expression of an adjacent or nearby protein-coding gene have been described. Here we examine the regulation of a locus consisting of two contiguous lncRNAs and the master regulator for entry into yeast meiosis, IME1. We find that the cluster of two lncRNAs together with several transcription factors form a regulatory circuit by which IME1 controls its own promoter and thereby promotes its own expression...
February 22, 2018: Nature Communications
Yuen-Ling Chan, Douglas K Bishop
Budding yeast Dmc1 is a member of the RecA family of strand exchange proteins essential for homologous recombination (HR) during meiosis. Dmc1 mediates the steps of homology search and DNA strand exchange reactions that are central to HR. To achieve optimum activity, Dmc1 requires a number of accessory factors. Although methods for purification of Dmc1 and many of its associated factors have been described (Binz, Dickson, Haring, & Wold, 2006; Busygina et al., 2013; Chan, Brown, Qin, Handa, & Bishop, 2014; Chi et al...
2018: Methods in Enzymology
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