mutant analysis
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2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Simone Höfler ◽  
Peer Lukat ◽  
Wulf Blankenfeldt ◽  
Teresa Carlomagno

AbstractBox C/D ribonucleoprotein complexes are RNA-guided methyltransferases that methylate the ribose 2’-OH of RNA. The central ‘guide RNA’ has box C and D motifs at its ends, which are crucial for activity. Archaeal guide RNAs have a second box C’/D’ motif pair that is also essential for function. This second motif is poorly conserved in eukaryotes and its function is uncertain. Conflicting literature data report that eukaryotic box C’/D’ motifs do or do not bind proteins specialized to recognize box C/D-motifs and are or are not important for function. Despite this uncertainty, the architecture of eukaryotic 2’-O-methylation enzymes is thought to be similar to that of their archaeal counterpart. Here, we use biochemistry, X-ray crystallography and mutant analysis to demonstrate the absence of functional box C’/D’ motifs in more than 80% of yeast guide RNAs. We conclude that eukaryotic Box C/D RNPs have two non-symmetric protein assembly sites and that their three-dimensional architecture differs from that of archaeal 2’-O-methylation enzymes.


Genetics ◽  
2021 ◽  
Author(s):  
Jacinta Davis ◽  
Claire Da Silva Santos ◽  
Narda Caudillo Zavala ◽  
Nicholas Gans ◽  
Daniel Patracuolla ◽  
...  

Abstract Parkinson’s Disease (PD) is primarily characterized by the loss of dopaminergic (DA) neurons in the brain. However, little is known about why DA neurons are selectively vulnerable to PD. To identify genes that are associated with DA neuron loss, we screened through 201 wild-caught populations of Drosophila melanogaster as part of the Drosophila Genetic Reference Panel (DGRP). Here we identify the top associated genes containing SNPs that render DA neurons vulnerable. These genes were further analyzed by using mutant analysis and tissue-specific knockdown for functional validation. We found that this loss of DA neurons caused progressive locomotor dysfunction in mutants and gene knockdown analysis. The identification of genes associated with the progressive loss of DA neurons should help to uncover factors that render these neurons vulnerable in PD, and possibly develop strategies to make these neurons more resilient.


Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 923
Author(s):  
Nicholas Cheung ◽  
Lei Tian ◽  
Xueru Liu ◽  
Xin Li

Botrytis cinerea is one of the most destructive fungal pathogens affecting numerous plant hosts, including many important crop species. As a molecularly under-studied organism, its genome was only sequenced at the beginning of this century and it was recently updated with improved gene annotation and completeness. In this review, we summarize key molecular studies on B. cinerea developmental and pathogenesis processes, specifically on genes studied comprehensively with mutant analysis. Analyses of these studies have unveiled key genes in the biological processes of this pathogen, including hyphal growth, sclerotial formation, conidiation, pathogenicity and melanization. In addition, our synthesis has uncovered gaps in the present knowledge regarding development and virulence mechanisms. We hope this review will serve to enhance the knowledge of the biological mechanisms behind this notorious fungal pathogen.


2020 ◽  
Vol 31 (19) ◽  
pp. 2107-2114 ◽  
Author(s):  
Paola Zambon ◽  
Saravanan Palani ◽  
Shekhar Sanjay Jadhav ◽  
Pananghat Gayathri ◽  
Mohan K. Balasubramanian

This work reveals an in vivo role for Myosin II in actin dynamics, potentially in its disassembly and turnover. The work uses double mutant analysis to arrive at this conclusion using the fission yeast as a model organism.


2020 ◽  
Author(s):  
Lohith Kunyeit ◽  
Nawneet K Kurrey ◽  
Anu-Appaiah K A ◽  
Reeta P Rao

AbstractBeneficial microbes in the intestine are thought to control pathogen overgrowth by competing for limited nutrients. Our findings modify this prevailing paradigm of a passive, microbial antagonistic mode of action to an active, directed mechanism mediated by specific secondary metabolites. We describe two food-derived yeasts, Saccharomyces cerevisiae and Issatchenkia occidentalis, that inhibit virulence traits of Candida albicans, including hyphal morphogenesis, biofilms formation and adhesion to intestinal epithelial cells. These yeasts also protect the model host Caenorhabditis elegans from C. albicans infection. We demonstrate that the protective activity is primarily retained in the secretome of the beneficial yeasts and the protection they provide as a physical barrier is minimal. Mutant analysis demonstrates that phenylethanol and tryptophol are necessary for protection and experiments with commercially procured compounds indicates that they are sufficient to inhibit C. albicans virulence. We propose food-derived yeasts as an alternative or combination therapy to conventional antifungal therapy for C. albicans infection.


2020 ◽  
Author(s):  
Lourds M. Fernando ◽  
Silwat Adeel ◽  
Mohammed Abul Basar ◽  
Anna K. Allen ◽  
Atanu Duttaroy

ABSTRACTThe nematode C. elegans has a contingent of five sod genes, one of the largest among aerobic organism. Earlier studies revealed each of the five sod genes is capable of making perfectly active SOD proteins in heterologous expressions systems therefore none appears to be a pseudogene. Yet deletion of the entire contingent of sod genes fails to impose any effect on the survival of C. elegans except these animals appear more sensitive to extraneously applied oxidative stress condition. We asked how many of the five sod genes are actually active in C. elegans through an in-gel SOD activity analysis. Here we provide evidence that out of the five genes only the mitochondrial SOD gene is active in C. elegans, albeit at a much lesser amount compared to D. melanogaster and E. coli. Mutant analysis further confirmed that among the mitochondrial forms, SOD-2 is the only naturally active SOD in C. elegans.


2020 ◽  
Vol 31 (12) ◽  
pp. 1218-1231 ◽  
Author(s):  
Kei Saito ◽  
Takashi Murayama ◽  
Tomone Hata ◽  
Takuya Kobayashi ◽  
Keitaro Shibata ◽  
...  

Using nanogold labeling and deletion mutant analysis, we determined the domain organization of dynactin subunit p150 and discovered that its CC1 domain adopted either a folded or an extended form. Furthermore, the entire sidearm of dynactin exhibited several characteristic forms, and the equilibrium among them depended on salt concentrations.


2020 ◽  
Vol 26 (5) ◽  
pp. 707-721.e5 ◽  
Author(s):  
Chengyi Sun ◽  
Hiromi Sakashita ◽  
Jang Kim ◽  
Zifeng Tang ◽  
G. Michael Upchurch ◽  
...  

2020 ◽  
Vol 226 (2) ◽  
pp. 541-554 ◽  
Author(s):  
Fengjiao Bu ◽  
Luuk Rutten ◽  
Yuda Purwana Roswanjaya ◽  
Olga Kulikova ◽  
Marta Rodriguez‐Franco ◽  
...  

2019 ◽  
Vol 375 (1792) ◽  
pp. 20190153 ◽  
Author(s):  
Susan K. Dutcher

The generation of ciliary waveforms requires the spatial and temporal regulation of dyneins. This review catalogues many of the asymmetric structures and proteins in the cilia of Chlamydomonas , a unicellular alga with two cilia that are used for motility in liquid medium. These asymmetries, which have been identified through mutant analysis, cryo-EM tomography and proteomics, provide a wealth of information to use for modelling how waveforms are generated and propagated. This article is part of the Theo Murphy meeting issue ‘Unity and diversity of cilia in locomotion and transport’.


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