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A mutation in budding yeast BRR6 affecting nuclear envelope insertion of the spindle pole body.

Jennifer M Gardner, Eileen O'Toole, Sue L Jaspersen,

<i>BRR6</i> and <i>BRL1</i> are two paralogs that encode transmembrane proteins of the nuclear envelope (NE) involved in membrane fluidity and nuclear pore complex biogenesis in organisms that undergo a closed mitosis. We show that mutation of a conserved cysteine in the intralumenal domain of <i>Saccharomyces cerevisiae</i> Brr6p results in a ... Read more >>

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[2021, 2021:]

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Improved CRISPR/Cas9 knock-in efficiency via the self-excising cassette (SEC) selection method in C. elegans.

George Huang, Bailey de Jesus, Alex Koh, Sara Blanco, Aubrie Rettmann, Ella DeMott, Melynda Sylvester, Cassie Ren, Carrie Meng, Skye Waterland, Anita Rhodes, Persephone Alicea, Abbey Flynn, Daniel J Dickinson, Ryan Doonan,

Streamlined, selection-based CRISPR knock-in protocols for <i>C. elegans</i> were first introduced six years ago (Dickinson <i>et al</i>. 2015; Schwartz and Jorgensen 2016). Though these selection-based approaches are powerful, one drawback has been the requirement to inject large numbers of P0 worms (~30-60 per gene target). We have found that a ... Read more >>

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[2021, 2021:]

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Decreased Mitochondrial Membrane Potential Activates the Mitochondrial Unfolded Protein Response.

Brandon J Berry, Tyrone O Nieves, Andrew P Wojtovich,

Mitochondria are ATP-producing organelles that also signal throughout the cell. Mitochondrial protein homeostasis is regulated through membrane potential-dependent protein import and quality control signaling. The mitochondrial unfolded protein response (UPR<sup>mt</sup>) is a specific program that responds to imbalances in nuclear and mitochondrial gene expression. Mounting evidence suggests that the electrochemical ... Read more >>

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[2021, 2021:]

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Modeling human glucose-6-phosphate dehydrogenase mutations using C. elegans GSPD-1.

Luiza N Loges, Katherine M Walstrom,

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked, recessive condition that causes intermittent jaundice or hemolytic anemia because of low NADPH levels in red blood cells. We performed steady-state enzyme kinetics with the recombinant <i>C. elegans</i> ortholog of human G6PD, GSPD-1, and two mutants containing amino acid changes found in human ... Read more >>

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[2021, 2021:]

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Drosophila insulin receptor regulates diabetes-induced mechanical nociceptive hypersensitivity.

Harika Dabbara, Arielle Schultz, Seol Hee Im,

Painful diabetic neuropathy (PDN) is one of the predominant complications of diabetes that causes numbness, tingling, and extreme pain sensitivity. Understanding the mechanisms of PDN pathogenesis is important for patient treatments. Here we report <i>Drosophila</i> models of diabetes-induced mechanical nociceptive hypersensitivity. Type 2 diabetes-like conditions and loss of insulin receptor ... Read more >>

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[2021, 2021:]

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Missense mutation of a conserved residue in UNC-112 (kindlin) eliminates binding to PAT-4 (ILK).

Hiroshi Qadota, Andres F Oberhauser, Guy M Benian,

<i>C. elegans</i> UNC-112 (kindlin) is required for muscle sarcomere assembly, and is one component of a conserved four-protein complex that associates with the cytoplasmic tail of integrin at the base of integrin adhesion complexes in muscle. The four-protein complex consists of UNC-112 (kindlin), PAT-4 (integrin linked kinase; ILK), PAT-6 (alpha-parvin), ... Read more >>

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[2021, 2021:]

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Meiotic H3K9me2 distribution is influenced by the ALG-3 and ALG-4 pathway and by poly(U) polymerase activity.

Yini Li, Matthew Snyder, Eleanor M Maine,

Histone modifications influence gene expression and chromosome dynamics by altering chromatin structure and recruitment of nonhistone proteins. Dimethylation of histone H3 on lysine 9 (H3K9me2) is a conserved modification often found within heterochromatin. During first meiotic prophase when homologous chromosomes undergo pairing and synapsis, immunolabeling of <i>C. elegans</i> male germ ... Read more >>

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[2021, 2021:]

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Transgenic reporter analysis of ChIP-Seq-defined enhancers identifies novel target genes for the terminal selector UNC-3/Collier/Ebf.

Yinan Li, Paschalis Kratsios,

Terminal selector-type transcription factors are key regulators of neuronal identity and function (Hobert and Kratsios, 2019; Kratsios and Hobert, 2018). Mechanistically, terminal selectors are thought to act directly through binding at the <i>cis</i>-regulatory region of genes (termed "terminal identity genes") that encode, among others, neurotransmitter [NT] synthesis proteins, ion channels, ... Read more >>

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[2021, 2021:]

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Deletion of a putative HDA-1 binding site in the hlh-2 promoter eliminates expression in C. elegans dorsal uterine cells.

Taylor N Medwig-Kinney, Nicholas J Palmisano, David Q Matus,

The helix-loop-helix transcription factor <i>hlh-2</i> (E/Daughterless) has been shown to play an important role in regulating cell fate patterning, cell cycle, and basement membrane invasion in the context of the development of the <i>C. elegans</i> somatic gonad. Here, using CRISPR/Cas9 genome engineering, we generated a new <i>hlh-2</i> allele (<i>hlh-2(Δ-1303-702)</i>) in ... Read more >>

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[2021, 2021:]

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The odd-1(tm848) mutation has no significant effect on brood size in Caenorhabditis elegans.

Gabriella M Scoca, Amy C Groth,

The genome of <i>Caenorhabditis elegans</i> contains two genes of the odd-skipped transcription factor family, <i>odd-1</i> and <i>odd-2.</i> In <i>C. elegans, odd-1</i> is expressed in the intestine. A deletion mutant (<i>tm848)</i> that removes most of the <i>odd-1</i> gene, including all three zinc fingers, has no significant effect on brood size when ... Read more >>

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[2021, 2021:]

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Improved GAL4 and Tet OFF drivers for C. elegans bipartite expression.

Michael Nonet,

The first generation of <i>C. elegans</i> GAL4 drivers for bipartite expression function less well than <i>C. elegans</i> tet ON/OFF, QF and LexA drivers. The main difference between the GAL4 drivers and the others is the absence of a flexible linker between the DNA binding and activation domain in the GAL4 ... Read more >>

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[2021, 2021:]

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Live-cell imaging reveals square shape spindles and long mitosis duration in the silkworm holocentric cells.

Lucien Vanpoperinghe, Frederique Carlier-Grynkorn, Gaetan Cornilleau, Takahiro Kusakabe, Ines A Drinnenberg, Phong T Tran,

Proper chromosome segregation during mitosis requires both the assembly of a microtubule (MT)-based spindle and the assembly of DNA-centromere-based kinetochore structure. Kinetochore-to-MT attachment enables chromosome separation. Monocentric cells, such as found in human, have one unique kinetochore per chromosome. Holocentric cells, such as found in the silkworm, in contrast, have ... Read more >>

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[2021, 2021:]

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ATG8 is conserved between Saccharomyces cerevisiae and psychrophilic, polar-collected fungi.

Brenna J Ivory, Hannah M Smith, Elizabeth Cabrera, Meaghan R Robinson, Jackson T Sparks, Amanda Solem, Jun-Ichi Ishihara, Hiroki Takahashi, Masaharu Tsuji, Verónica A Segarra,

Autophagy is a conserved catabolic process by which eukaryotic cells respond to stress by targeting damaged or unneeded molecules or organelles for sequestration into specialized vesicles known as autophagosomes. Autophagosomes ultimately facilitate the digestion and recycling of their contents by fusing with the degradative organelle of the cell. Studies of ... Read more >>

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[2021, 2021:]

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The Caenorhabditis elegans tmc-1 is involved in egg-laying inhibition in response to harsh touch.

Eva Kaulich, Denise S Walker, Yi-Quan Tang, William Ronald Schafer,

The conserved family of Transmembrane channel-like (TMC) proteins has attracted significant interest since two members appear to be key components of the mammalian hair cell mechanotransducer involved in hearing. <i>C. elegans</i> expresses two TMC proteins, TMC-1 and TMC-2. TMC-1 is widely expressed in in both muscles and the nervous system. ... Read more >>

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[2021, 2021:]

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Wild Caught Nematode Identification and Early Embryo Development: An accessible undergraduate research experience.

Haley Attix, Alex George, Henali Panchal, Angel Cortez, Martin Cho, Kathy Zarilla, Eric Hastie,

Ample evidence suggests that participation in undergraduate research in community college is critical for stimulating interest and retention in STEM careers. Guided skill development and practice in a collaborative lab setting allows students to be competitive when applying to future research opportunities. The goals of this undergraduate research experience (URE) ... Read more >>

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[2021, 2021:]

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Direct comparisons of bisulfite pyrosequencing versus targeted bisulfite sequencing.

Dillon E King, A Clare Sparling, Rashmi Joglekar, Joel N Meyer, Susan K Murphy,

DNA methylation is an important epigenetic mechanism involved in proper genome function. Bisulfite pyrosequencing (PSQ) is a commonly used technique to quantify DNA methylation. Although very accurate, bisulfite pyrosequencing can be expensive and time consuming for large-scale quantitative DNA methylation analysis at the single nucleotide level. High throughput DNA methylation ... Read more >>

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[2021, 2021:]

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Deficient mechanosensation in mec-3 decreases precipice response in C. elegans.

Robin M Mitchell, Diana S Pattillos, Shuyu Zhang, Jared J Young,

The precipice response in <i>Caenorhabditis elegans</i> is a little-understood phenomenon in which worms move rapidly away from edges. We hypothesized that mechanosensation underlies the precipice response and that mechanosensory mutants would exhibit the precipice response less often than N2 wild type worms. We found that <i>mec-3</i> mutants, with severe loss ... Read more >>

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[2021, 2021:]

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GPN-1/glypican and UNC-52/perlecan do not appear to function in BMP signaling to pattern the C. elegans postembryonic mesoderm.

Melisa S DeGroot, Robert Greer, Jun Liu,

Heparan sulfate proteoglycans (HSPGs) are diverse macromolecules consisting of a protein core modified with glycosaminoglycan (GAG) chains. HSPGs, including glypicans and perlecans, have been implicated in shaping the extracellular matrix (ECM) to affect growth factor signaling. Here, we tested if GPN-1/glypicanor UNC-52/perlecan plays a role in the bone morphogenetic protein ... Read more >>

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[2021, 2021:]

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Sorbitol and PKC1 overexpression alleviate temperature sensitivity in chaperonin mutants of Saccharomyces cerevisiae.

Aswathy Narayanan, M Anaul Kabir,

CCT (Chaperonin containing TCP-1) is a constitutively expressed eukaryotic chaperonin complex involved in the proper folding of proteins like actin and tubulin. Temperature sensitive mutants of CCT complex have been employed in various genetic screens, acting as models to study human CCT, the defects of which are implicated in disease ... Read more >>

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[2021, 2021:]

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Suppression of him-14(it44ts) by a transgene insertion expressing GFP::COSA-1.

Chloé Girard, Chantal C Akerib, Anne M Villeneuve,

Meiotic crossover formation requires the activity of multiple pro-crossover factors, including the MutSγ complex and the cyclin-related protein COSA-1, that become concentrated together at the sites of crossover recombination intermediates. Here we show that a transgene insertion expressing GFP::COSA-1 can suppress the crossover deficit caused by a partial reduction in ... Read more >>

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[2021, 2021:]

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EGL-4 promotes turning behavior of C. elegans males during mating.

Shapour Rahmani, Simon Tuck,

During mating, <i>C. elegans</i> males whose tails have reached the head or tail of their intended mates are able to switch to scanning the other side by performing a turn during which the male's tail curls ventrally all the while keeping in contact with the hermaphrodite. The ability to execute ... Read more >>

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[2021, 2021:]

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Widespread gpa-4 promoter-driven expression during Caenorhabditis elegans development.

Jaime Osuna-Luque, Collin Y Ewald, Peter Meister,

The <i>gpa-4</i> promoter-driven expression is described as specific for ASIL and ASIR chemosensory neurons in the nematode <i>Caenorhabditis elegans</i>, yet this was mostly examined in adult animals. Here we used a recombination-mediated reporter to test the previously used <i>gpa-4</i> promoter expression. This reporter highlights all cells in which the <i>gpa-4</i> ... Read more >>

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[2021, 2021:]

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Two plasmid modules for introducing the auxin-inducible degron into the fission yeast Schizosaccharomyces pombe by PCR-based gene targeting.

Xiuyi Song, Ruoming Xu, Tomoyasu Sugiyama,

Targeted protein degradation is a powerful approach to study and inhibit protein function <i>in vivo</i>. Introduction of the auxin-inducible degron (AID) system to the fission yeast <i>Schizosaccharomyces pombe</i> was previously reported, but, to the best of our knowledge, no plasmid for constructing AID-tagged fission yeast strains has been described so ... Read more >>

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[2021, 2021:]

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The primordial germ line is refractory to perturbations of actomyosin regulator function in C. elegans L1 larvae.

Jack Bauer, Léa Lacroix, Jean-Claude Labbé,

Cytokinesis, the separation of daughter cells at the end of mitosis, relies on the coordinated activity of several regulators of actomyosin assembly and contractility (Green et al. 2012). These include the small GTPase RhoA (RHO-1) and its guanine-nucleotide exchange factor Ect2 (ECT-2), the scaffold protein Anillin (ANI-1), the non-muscle myosin ... Read more >>

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[2021, 2021:]

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DMSO reduces the cytotoxicity of anticancer ruthenium complex KP1019 in yeast.

Jonathan Davis, Anne Cetto, Mary Campbell, Seth Scoggins, Laura Stultz, Pamela Hanson,

Low solubility in aqueous solutions is a significant limitation of the otherwise promising anticancer ruthenium complex KP1019. In laboratory studies, this challenge is often overcome by using DMSO to help drive the drug into solution. Since DMSO was previously shown to alter the bioactivity of platinum-based chemotherapeutics, here we examine ... Read more >>

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[2021, 2021:]

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