Reduced dosage of multiple dynein arm preassembly factors limits cilia regeneration inChlamydomonas reinhardtii DOI Creative Commons
Gervette M. Penny, Susan K. Dutcher

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Ноя. 2, 2023

Abstract Motile cilia are complex organelles comprised of >800 structural proteins. Variants in 58 these genes cause primary ciliary dyskinesia (PCD) humans. We used a second-site non-complementation approach diploid Chlamydomonas reinhardtii strains to investigate whether reduced dosage different combinations motile proteins affects growth / regeneration. Temperature-sensitive mutants intraflagellar transport fail complement heterozygous diploids at the restrictive temperature, which may be due poisonous interactions or reduction protein levels. Diploid for 211 double mutant 21 phenotypically recessive when unperturbed and do not show SSNC. Consequently, we developed sensitized screen. When synthesis is inhibited, cells utilize existing cytoplasmic pool proteins, insufficient build full-length cilia. Six dynein arm preassembly factors (DNAAFs) pf13, pf23, wdr92, oda8 regrow that shorter than wild-type diploids, suggests heterozygosity limits assembled arms further. isolated new null allele ( pf23-4) shows more severe loss ODAs IDAs previously studied pf23-1 hypomorphic allele. also identified role PF23 modification IC138, I1/ f inner arm. In our SSNC assay, pf23-4 exhibits phenotype . Whole cell extract immunoblots one-half levels both single genotypes. The pf13 with two outer ODA6 , ODA9. suggest multiple needed assembly Our data support as scaffolding hub involved assembly. Author Summary essential movement fluids. humans, defects (PCD), rare disease characterized by recurring respiratory infections, left-right asymmetry defects, ear infertility. Many function use study effects changes results help fold assemble motors cytoplasm sensitive factors. Reductions steps pathway prevents efficient regeneration under stressful cellular conditions.

Язык: Английский

Near telomere-to-telomere genome of the model plant Physcomitrium patens DOI
Guiqi Bi,

Shijun Zhao,

Jiawei Yao

и другие.

Nature Plants, Год журнала: 2024, Номер 10(2), С. 327 - 343

Опубликована: Янв. 26, 2024

Язык: Английский

Процитировано

39
Technology-enabled great leap in deciphering plant genomes DOI
Lingjuan Xie, Xiaojiao Gong, Kun Yang

и другие.

Nature Plants, Год журнала: 2024, Номер 10(4), С. 551 - 566

Опубликована: Март 20, 2024

Язык: Английский

Процитировано

39
The Chlamydomonas Genome Project, version 6: Reference assemblies for mating-type plus and minus strains reveal extensive structural mutation in the laboratory DOI Creative Commons
Rory J. Craig, Sean D. Gallaher, Shengqiang Shu

и другие.

The Plant Cell, Год журнала: 2022, Номер 35(2), С. 644 - 672

Опубликована: Дек. 23, 2022

Abstract Five versions of the Chlamydomonas reinhardtii reference genome have been produced over last two decades. Here we present version 6, bringing significant advances in assembly quality and structural annotations. PacBio-based chromosome-level assemblies for laboratory strains, CC-503 CC-4532, provide resources plus minus mating-type alleles. We corrected major misassemblies previous validated our via linkage analyses. Contiguity increased ten-fold >80% filled gaps are within genes. used Iso-Seq deep RNA-seq datasets to improve annotations, updated gene symbols textual annotation functionally characterized genes extensive manual curation. discovered that cell wall-less classical strain exhibits genomic instability potentially caused by deletion helicase RECQ3, with mutations identified affect >100 therefore CC-4532 as primary reference, although this also carries unique is experiencing rapid proliferation a Gypsy retrotransposon. expect all strains harbor gene-disrupting mutations, which should be considered when interpreting comparing experimental results. Collectively, presented here herald new era genomics will foundation continued research important organism.

Язык: Английский

Процитировано

62
Cryptic infection of a giant virus in a unicellular green alga DOI
Maria P. Erazo-Garcia, Uri Sheyn, Zachary K. Barth

и другие.

Science, Год журнала: 2025, Номер unknown

Опубликована: Апрель 10, 2025

Latency is a common strategy in wide range of viral lineages, but its prevalence giant viruses remains unknown. Here we describe 617 kbp integrated element the model green alga Chlamydomonas reinhardtii . We resolve genome using long-read sequencing, identify putative polinton-like integrase, and show that particles accumulate primarily during stationary growth phase. A diverse array viral-encoded selfish genetic elements expressed activity, including several Fanzor nuclease-encoding transposable elements. In addition, field isolates sp. harbor signatures endogenous related to C. virus exhibit similar infection dynamics, suggesting latency prevalent natural host communities. Our work describes an unusually large temperate unicellular eukaryote, substantially expanding scope cryptic infections virosphere.

Язык: Английский

Процитировано

1
Protofilament-specific nanopatterns of tubulin post-translational modifications regulate the mechanics of ciliary beating DOI Creative Commons
Gonzalo Alvarez Viar, Nikolai Klena,

Fabrizio Martino

и другие.

Current Biology, Год журнала: 2024, Номер unknown

Опубликована: Сен. 1, 2024

Язык: Английский

Процитировано

7
Gene dosage of independent dynein arm motor preassembly factors influences cilia assembly in Chlamydomonas reinhardtii DOI Creative Commons
Gervette M. Penny, Susan K. Dutcher

PLoS Genetics, Год журнала: 2024, Номер 20(3), С. e1011038 - e1011038

Опубликована: Март 18, 2024

Motile cilia assembly utilizes over 800 structural and cytoplasmic proteins. Variants in approximately 58 genes cause primary ciliary dyskinesia (PCD) humans, including the dynein arm (pre)assembly factor (DNAAF) gene DNAAF4 . In outer arms (ODAs) inner (IDAs) fail to assemble motile when function is disrupted. Chlamydomonas reinhardtii , a ciliated unicellular alga, ortholog called PF23 The pf23-1 mutant assembles short lacks IDAs, but partially retains ODAs. of new null allele ( pf23-4 ) completely lack ODAs IDAs are even shorter than from addition, plays role modification IC138, protein two-headed IDA (I1/f). As most PCD variants humans recessive, we sought test if heterozygosity at two affects using second-site non-complementation (SSNC) screening approach. We asked phenotypes were observed diploids with pairwise heterozygous combinations 21 well-characterized strains. Vegetative cultures single double diploid cells did not show SSNC for motility phenotypes. When synthesis inhibited, wild-type utilize pool proteins half-length cilia. this sensitized assay, 8 pf23 other DNAAF mutations SSNC; they wild-type. contrast, 203 strains showed no effect on assembly. Immunoblots wdr92 or oda8 that reduced by half these strains, dosage phenotype severity. Reductions another affect ability impedes Thus, multiple DNAAFs an important regeneration.

Язык: Английский

Процитировано

5
Chlamydomonas FBB18 is a ubiquitin-like protein essential for the cytoplasmic preassembly of various ciliary dyneins DOI Creative Commons
Ryosuke Yamamoto,

Yui Sahashi,

Rieko Shimo‐Kon

и другие.

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(12)

Опубликована: Март 19, 2025

Motile cilia are organelles found on many eukaryotic cells that play critical roles in development and fertility. Human CFAP298 has been implicated the transport/assembly of ciliary dyneins, defects this protein cause primary dyskinesia. However, neither exact function nor structure have elucidated. Here, we took advantage Chlamydomonas, a ciliated alga, to study FBB18, an ortholog CFAP298. Multiple dyneins were greatly reduced Chlamydomonas fbb18 mutants. In addition, both stability dynein heavy chains (HCs) association between HCs intermediate/light (IC/LCs) cytoplasm, strongly suggesting FBB18 functions cytoplasmic assembly (the so-called "preassembly") complexes from HC/IC/LCs. Furthermore, X-ray crystallography revealed forms bilobed with globular domains at ends molecule, connected by α-helical bundle. Unexpectedly, one domain shows high similarity ubiquitin, small for modification variety complexes, ubiquitin-like is indispensable molecular FBB18. Our results demonstrate specialized member family, plays role preassembly, most likely mediating diverse interactions HCs, chaperone(s), other preassembly factor(s) using as well regions, facilitating proper folding HCs.

Язык: Английский

Процитировано

0
Elevated accumulation of lutein and zeaxanthin in a novel high-biomass yielding strain Dunaliella sp. ZP-1 obtained through EMS mutagenesis DOI Creative Commons
Chenglong Liu,

Danqiong Huang,

Xinran Zhuo

и другие.

Biotechnology for Biofuels and Bioproducts, Год журнала: 2025, Номер 18(1)

Опубликована: Март 27, 2025

Abstract Background Dunaliella microalgae, such as salina riching in β-carotene and bardawil rich lutein α-carotene, have been used aquaculture, supplements, cosmetics, feed industries. The genus is diverse; therefore, characterization of novel strains isolation new varieties through mutagenesis technology will promote natural carotenoid bioproduction. Results Salt stress test demonstrated that the newly isolated microalgae strain ZP-1 was a halotolerant strain. Morphology observation molecular phylogeny analysis indicated unicellular green microalga member . Biomass RAM medium up to 2.45 g/L, showing advantage over other common terms yield. Furthermore, Ethyl methanesulfonate (EMS) mutant library generated from this high-biomass strain, aiming improve productivity. A selected morphology combining with quantification by HPLC, which nominated turn yellow dunaliella 4 ( tyd4 ). displayed an increased productivity 28.55% zeaxanthin 22.19%. promoted 17.40% continuous culture under red light. Application exogenous 1.0 μM melatonin on led cell density improved biomass. Conclusions study support EMS effective breeding approach for further improvement sp. ZP-1, exhibiting potential overcome bottleneck low biomass current commercial strains. had higher contents both zeaxanthin, whose yield could be elevated light melatonin. This provided sources scientific research technical reference bioproduction carotenoids.

Язык: Английский

Процитировано

0
Genome editing in the green alga Chlamydomonas: past, present practice and future prospects DOI Creative Commons
Adrian P. Nievergelt

The Plant Journal, Год журнала: 2025, Номер 122(1)

Опубликована: Апрель 1, 2025

SUMMARY The green alga Chlamydomonas is an important and versatile model organism for research topics ranging from photosynthesis metabolism, cilia, basal bodies to cellular communication the cycle of significant interest bioengineering processes. genome in this unicellular contained 17 haploid chromosomes codes 16 883 protein coding genes. Functional genomics, as well biotechnological applications, rely on ability remove, add, change these genes a controlled efficient manner. In review, history gene editing put context wider developments genetics demonstrate how many key engineer algae follow global trends availability technology. Building background, overview state art engineering given, focusing primarily practical aspects while giving examples recent applications. Commonly encountered ‐specific challenges, developments, community resources are presented, finally, comprehensive discussion emergence evolution CRISPR/Cas‐based precision given. An outline possible future paths based current genetic tools presented.

Язык: Английский

Процитировано

0
Internal ploidy and heritable chromosome fragmentation in Parachlorella kessleri DOI Creative Commons

Kotaro Ishii,

Michiko Asano, Yusuke Kazama

и другие.

Algal Research, Год журнала: 2025, Номер unknown, С. 104047 - 104047

Опубликована: Апрель 1, 2025

Язык: Английский

Процитировано

0