title: A Forward Genetic Screen and Whole Genome Sequencing Identify Deflagellation Defective Mutants in Chlamydomonas, Including Assignment of ADF1 as a TRP Channel.
abstract: 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.
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Laura K
authors: 0000-0002-6413-6586
authors: LK
authors: Hilton
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Fabian
authors: F
authors: Meili
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Paul D
authors: PD
authors: Buckoll
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Julie C
authors: JC
authors: Rodriguez-Pike
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Courtney P
authors: CP
authors: Choutka
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Jaime A
authors: JA
authors: Kirschner
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Freda
authors: F
authors: Warner
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Mette
authors: M
authors: Lethan
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Fabian A
authors: FA
authors: Garces
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada.
authors: Jingnan
authors: J
authors: Qi
authors: Department of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, British Columbia, V5A 1S6, Canada quarmby@sfu.ca.
authors: Lynne M
authors: 0000-0001-6661-680X
authors: LM
authors: Quarmby
mesh_terms: D002696:Chlamydomonas / Q000235:genetics*; D016825:Chlamydomonas reinhardtii / Q000235:genetics; D002874:Chromosome Mapping; D004252:DNA Mutational Analysis; D005407:Flagella / Q000235:genetics*; D023061:Gene Order; D005820:Genetic Testing; D018745:Genome, Plant; D023281:Genomics / Q000379:methods; D059014:High-Throughput Nucleotide Sequencing; D009154:Mutation; D010802:Phylogeny; D011995:Recombination, Genetic; D050051:Transient Receptor Potential Channels / Q000145:classification / Q000235:genetics*
chemical_list: D050051:Transient Receptor Potential Channels
keywords: Chlamydomonas reinhardtii; FAP16; Mutant Screen Report; TRP15; calcium