Dataset Information

A large-scale genetic screen identifies genes essential for motility in Agrobacterium fabrum.

ABSTRACT: The genetic and molecular basis of flagellar motility has been investigated for several decades, with innovative research strategies propelling advances at a steady pace. Furthermore, as the phenomenon is examined in diverse bacteria, new taxon-specific regulatory and structural features are being elucidated. Motility is also a straightforward bacterial phenotype that can allow undergraduate researchers to explore the palette of molecular genetic tools available to microbiologists. This study, driven primarily by undergraduate researchers, evaluated hundreds of flagellar motility mutants in the Gram-negative plant-associated bacterium Agrobacterium fabrum. The nearly saturating screen implicates a total of 37 genes in flagellar biosynthesis, including genes of previously unknown function.

SUBMITTER: Calvopina-Chavez DG

PROVIDER: S-EPMC9812332 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature

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A large-scale genetic screen identifies genes essential for motility in Agrobacterium fabrum.

Calvopina-Chavez Diana G DG Howarth Robyn E RE Memmott Audrey K AK Pech Gonzalez Oscar H OH Hafen Caleb B CB Jensen Kyson T KT Benedict Alex B AB Altman Jessica D JD Burnside Brittany S BS Childs Justin S JS Dallon Samuel W SW DeMarco Alexa C AC Flindt Kirsten C KC Grover Sarah A SA Heninger Elizabeth E Iverson Christina S CS Johnson Abigail K AK Lopez Jack B JB Meinzer McKay A MA Moulder Brook A BA Moulton Rebecca I RI Russell Hyrum S HS Scott Tiana M TM Shiobara Yuka Y Taylor Mason D MD Tippets Kathryn E KE Vainerere Kayla M KM Von Wallwitz Isabella C IC Wagley Madison M Wiley Megumi S MS Young Naomi J NJ Griffitts Joel S JS

PloS one 20230104 1

The genetic and molecular basis of flagellar motility has been investigated for several decades, with innovative research strategies propelling advances at a steady pace. Furthermore, as the phenomenon is examined in diverse bacteria, new taxon-specific regulatory and structural features are being elucidated. Motility is also a straightforward bacterial phenotype that can allow undergraduate researchers to explore the palette of molecular genetic tools available to microbiologists. This study, d ...[more]

PMID: 36598925

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Project description:Neuronal circuit development and function require proper synapse formation and maintenance. Genetic screens are one powerful method to identify the mechanisms shaping synaptic development and stability. However, genes with essential roles in non-neural tissues may be missed in traditional loss-of-function screens. In an effort to circumvent this limitation, we used neuron-specific RNAi knock down in Drosophila and assayed the formation, growth, and maintenance of the neuromuscular junction (NMJ). We examined 1970 Drosophila genes, each of which has a conserved ortholog in mammalian genomes. Knock down of 158 genes in post-mitotic neurons led to abnormalities in the neuromuscular system, including misapposition of active zone components opposite postsynaptic glutamate receptors, synaptic terminal overgrowth and undergrowth, abnormal accumulation of synaptic material within the axon, and retraction of synaptic terminals from their postsynaptic targets. Bioinformatics analysis demonstrates that genes with overlapping annotated function are enriched within the hits for each phenotype, suggesting that the shared biological function is important for that aspect of synaptic development. For example, genes for proteasome subunits and mitotic spindle organizers are enriched among the genes whose knock down leads to defects in synaptic apposition and NMJ stability. Such genes play essential roles in all cells, however the use of tissue- and temporally-restricted RNAi indicates that the proteasome and mitotic spindle organizers participate in discrete aspects of synaptic development. In addition to identifying functional classes of genes shaping synaptic development, this screen also identifies candidate genes whose role at the synapse can be validated by traditional loss-of-function analysis. We present one such example, the dynein-interacting protein NudE, and demonstrate that it is required for proper axonal transport and synaptic maintenance. Thus, this screen has identified both functional classes of genes as well as individual candidate genes that are critical for synaptic development and will be a useful resource for subsequent mechanistic analysis of synapse formation and maintenance.

Dataset Information

A large-scale genetic screen identifies genes essential for motility in Agrobacterium fabrum.

Publications

A large-scale genetic screen identifies genes essential for motility in Agrobacterium fabrum.

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