Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein

Autor: Magdalena Bezanilla, Xinxin Ding, Carl Bascom, Allison M. Butt, Sakshi Khurana, Luis Vidali, Patrick Flaherty, Jeffrey P. Bibeau, Leah M. Pervere, Robert G. Orr
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Cancer Research
Cell division
Mutant
Artificial Gene Amplification and Extension
Plant Science
Plant Genetics
medicine.disease_cause
Polymerase Chain Reaction
Microtubules
Database and Informatics Methods
Sequencing techniques
Gene Expression Regulation
Plant
Plant Genomics
DNA sequencing
Nonvascular Plants
Cytoskeleton
Flowering Plants
Genetics (clinical)
Plant Proteins
Mutation
biology
Chromosome Mapping
Eukaryota
Genomics
Plants
Cell biology
Phenotypes
Phenotype
Cell Processes
Perspective
RNA Interference
Cellular Structures and Organelles
Sequence Analysis
Transcriptome Analysis
Research Article
Biotechnology
Next-Generation Sequencing
lcsh:QH426-470
Bioinformatics
Microtubule Polymerization
Microtubule Dynamics
Research and Analysis Methods
Physcomitrella patens
Chromosomes
Plant
03 medical and health sciences
Microtubule
Mosses
Genetics
medicine
Point Mutation
Molecular Biology Techniques
Molecular Biology
Gene
Ecology
Evolution
Behavior and Systematics
Whole Genome Sequencing
Gene Mapping
Organisms
Biology and Life Sciences
Computational Biology
Cell Biology
Genome Analysis
biology.organism_classification
Bryopsida
lcsh:Genetics
030104 developmental biology
Plant Biotechnology
Homologous recombination
Sequence Alignment
Genetic screen
Zdroj: PLoS Genetics, Vol 14, Iss 5, p e1007221 (2018)
PLoS Genetics
ISSN: 1553-7404
1553-7390
Popis: Our ability to identify genes that participate in cell growth and division is limited because their loss often leads to lethality. A solution to this is to isolate conditional mutants where the phenotype is visible under restrictive conditions. Here, we capitalize on the haploid growth-phase of the moss Physcomitrella patens to identify conditional loss-of-growth (CLoG) mutants with impaired growth at high temperature. We used whole-genome sequencing of pooled segregants to pinpoint the lesion of one of these mutants (clog1) and validated the identified mutation by rescuing the conditional phenotype by homologous recombination. We found that CLoG1 is a novel and ancient gene conserved in plants. At the restrictive temperature, clog1 plants have smaller cells but can complete cell division, indicating an important role of CLoG1 in cell growth, but not an essential role in cell division. Fluorescent protein fusions of CLoG1 indicate it is localized to microtubules with a bias towards depolymerizing microtubule ends. Silencing CLoG1 decreases microtubule dynamics, suggesting that CLoG1 plays a critical role in regulating microtubule dynamics. By discovering a novel gene critical for plant growth, our work demonstrates that P. patens is an excellent genetic system to study genes with a fundamental role in plant cell growth.
Author summary Genes important for cell growth are difficult to identify because their disruption often results in the death of the organism. A solution to this problem is to isolate temperature-sensitive mutants where growth is blocked only at high temperatures. Here, we used the moss Physcomitrella patens, a simple model plant, to isolate temperature-sensitive mutants with reduced growth. We used whole-genome sequencing to identify the gene disrupted in one of these mutants (clog1). We found that CLoG1 is a previously uncharacterized gene present in algae and plants. Localization studies of CLoG1 protein in living cells showed CLoG1 concentrates on microtubules and tracks depolymerizing ends. Loss-of-function analysis suggests a possible role in controlling microtubule dynamics. Our approach establishes the moss P. patens as a valuable model-organism to investigate genes important for cell growth in plants.
Databáze: OpenAIRE
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