A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel
With rare exception, ciliated cells entering mitosis lose their cilia, thereby freeing basal bodies to serve as centrosomes in the formation of high-fidelity mitotic spindles. Cilia can be lost by shedding or disassembly, but either way, it appears that the final release may be via a coordinated sev...
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2016-10-01
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Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.116.034264 |
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doaj-fea7afcf3d044fd2ab1b1283d62fa72f2021-07-02T03:12:03ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362016-10-016103409341810.1534/g3.116.03426435A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP ChannelLaura K. HiltonFabian MeiliPaul D. BuckollJulie C. Rodriguez-PikeCourtney P. ChoutkaJaime A. KirschnerFreda WarnerMette LethanFabian A. GarcesJingnan QiLynne M. QuarmbyWith rare exception, ciliated cells entering mitosis lose their cilia, thereby freeing basal bodies to serve as centrosomes in the formation of high-fidelity mitotic spindles. Cilia can be lost by shedding or disassembly, but either way, it appears that the final release may be via a coordinated severing of the nine axonemal outer doublet microtubules linking the basal body to the ciliary transition zone. Little is known about the mechanism or regulation of this important process. The stress-induced deflagellation response of Chlamydomonas provides a basis to identifying key players in axonemal severing. In an earlier screen we uncovered multiple alleles for each of three deflagellation genes, ADF1, FA1, and FA2. Products of the two FA genes localize to the site of axonemal severing and encode a scaffolding protein and a member of the NIMA-related family of ciliary-cell cycle kinases. The identity of the ADF1 gene remained elusive. Here, we report a new screen using a mutagenesis that yields point mutations in Chlamydomonas, an enhanced screening methodology, and whole genome sequencing. We isolated numerous new alleles of the three known genes, and one or two alleles each of at least four new genes. We identify ADF1 as a TRP ion channel, which we suggest may reside at the flagellar transition zone.http://g3journal.org/lookup/doi/10.1534/g3.116.034264calciumChlamydomonas reinhardtiiTRP15FAP16Mutant Screen Report |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Laura K. Hilton Fabian Meili Paul D. Buckoll Julie C. Rodriguez-Pike Courtney P. Choutka Jaime A. Kirschner Freda Warner Mette Lethan Fabian A. Garces Jingnan Qi Lynne M. Quarmby |
spellingShingle |
Laura K. Hilton Fabian Meili Paul D. Buckoll Julie C. Rodriguez-Pike Courtney P. Choutka Jaime A. Kirschner Freda Warner Mette Lethan Fabian A. Garces Jingnan Qi Lynne M. Quarmby A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel G3: Genes, Genomes, Genetics calcium Chlamydomonas reinhardtii TRP15 FAP16 Mutant Screen Report |
author_facet |
Laura K. Hilton Fabian Meili Paul D. Buckoll Julie C. Rodriguez-Pike Courtney P. Choutka Jaime A. Kirschner Freda Warner Mette Lethan Fabian A. Garces Jingnan Qi Lynne M. Quarmby |
author_sort |
Laura K. Hilton |
title |
A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel |
title_short |
A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel |
title_full |
A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel |
title_fullStr |
A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel |
title_full_unstemmed |
A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel |
title_sort |
forward genetic screen and whole genome sequencing identify deflagellation defective mutants in chlamydomonas, including assignment of adf1 as a trp channel |
publisher |
Oxford University Press |
series |
G3: Genes, Genomes, Genetics |
issn |
2160-1836 |
publishDate |
2016-10-01 |
description |
With rare exception, ciliated cells entering mitosis lose their cilia, thereby freeing basal bodies to serve as centrosomes in the formation of high-fidelity mitotic spindles. Cilia can be lost by shedding or disassembly, but either way, it appears that the final release may be via a coordinated severing of the nine axonemal outer doublet microtubules linking the basal body to the ciliary transition zone. Little is known about the mechanism or regulation of this important process. The stress-induced deflagellation response of Chlamydomonas provides a basis to identifying key players in axonemal severing. In an earlier screen we uncovered multiple alleles for each of three deflagellation genes, ADF1, FA1, and FA2. Products of the two FA genes localize to the site of axonemal severing and encode a scaffolding protein and a member of the NIMA-related family of ciliary-cell cycle kinases. The identity of the ADF1 gene remained elusive. Here, we report a new screen using a mutagenesis that yields point mutations in Chlamydomonas, an enhanced screening methodology, and whole genome sequencing. We isolated numerous new alleles of the three known genes, and one or two alleles each of at least four new genes. We identify ADF1 as a TRP ion channel, which we suggest may reside at the flagellar transition zone. |
topic |
calcium Chlamydomonas reinhardtii TRP15 FAP16 Mutant Screen Report |
url |
http://g3journal.org/lookup/doi/10.1534/g3.116.034264 |
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