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

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...

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Main Authors: Xinxin Ding, Leah M Pervere, Carl Bascom, Jeffrey P Bibeau, Sakshi Khurana, Allison M Butt, Robert G Orr, Patrick J Flaherty, Magdalena Bezanilla, Luis Vidali
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2018-05-01
Series:PLoS Genetics
Online Access:http://europepmc.org/articles/PMC5944918?pdf=render
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spelling doaj-900d78187fb64643be74f77d763779b82020-11-24T21:49:54ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042018-05-01145e100722110.1371/journal.pgen.1007221Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.Xinxin DingLeah M PervereCarl BascomJeffrey P BibeauSakshi KhuranaAllison M ButtRobert G OrrPatrick J FlahertyMagdalena BezanillaLuis VidaliOur 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.http://europepmc.org/articles/PMC5944918?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Xinxin Ding
Leah M Pervere
Carl Bascom
Jeffrey P Bibeau
Sakshi Khurana
Allison M Butt
Robert G Orr
Patrick J Flaherty
Magdalena Bezanilla
Luis Vidali
spellingShingle Xinxin Ding
Leah M Pervere
Carl Bascom
Jeffrey P Bibeau
Sakshi Khurana
Allison M Butt
Robert G Orr
Patrick J Flaherty
Magdalena Bezanilla
Luis Vidali
Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
PLoS Genetics
author_facet Xinxin Ding
Leah M Pervere
Carl Bascom
Jeffrey P Bibeau
Sakshi Khurana
Allison M Butt
Robert G Orr
Patrick J Flaherty
Magdalena Bezanilla
Luis Vidali
author_sort Xinxin Ding
title Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
title_short Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
title_full Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
title_fullStr Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
title_full_unstemmed Conditional genetic screen in Physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
title_sort conditional genetic screen in physcomitrella patens reveals a novel microtubule depolymerizing-end-tracking protein.
publisher Public Library of Science (PLoS)
series PLoS Genetics
issn 1553-7390
1553-7404
publishDate 2018-05-01
description 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.
url http://europepmc.org/articles/PMC5944918?pdf=render
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