Biology subjects

Fakhimi, N.

Publications and source records attributed to Fakhimi, N..

6 recordsLinked to original sources

bioRxiv2026

Dual-targeted NADK2 Links Mitochondrial Redox Homeostasis to Carbon Partitioning and Heterotrophic Growth in Chlamydomonas reinhardtii

NAD kinases (NADKs) can modulate NADP(H) levels across shifting environmental conditions. In Chlamydomonas reinhardtii, we localized CreNADK1 to the chloroplast, CreNADK3 to the cytoplasm, and CreNADK2 to both the chloroplast and mitochondria. Importantly, during mixotrophic (light + acetate) and photoautotrophic (light) growth, most CreNADK2 is chloroplast localized whereas an increased proportion resides in mitochondria when the cells are grown heterotrophically (dark + acetate). In Crenadk2 mutants, a diminished ability to synthesize mitochondrial NADP(H) inhibits the TCA cycle and markedly retards heterotrophic growth. In the light, photosynthesis becomes the primary energy source, and the presence of chloroplastic CreNADK1 enables near-normal growth in mutant cells, with inhibition of TCA cycle activity partially ameliorated via rerouting of acetate metabolism through the glyoxylate shunt, bypassing the TCA cycle and diminishing mitochondrial ROS production. These findings demonstrate spatial plasticity of NAD(H)Ks and the implementation of bypass metabolism to enable growth of Crenadk2 mutants.

cell biology↗

bioRxiv2023

Chloroplast Methyltransferase Homolog RMT2 is Involved in Photosystem I Biogenesis

Oxygen (O2), a dominant element in the atmosphere and essential for most life on Earth, is produced by the photosynthetic oxidation of water. However, metabolic activity can cause accumulation of reactive O2 species (ROS) and severe cell damage. To identify and characterize mechanisms enabling cells to cope with ROS, we performed a high-throughput O2 sensitivity screen on a genome-wide insertional mutant library of the unicellular alga Chlamydomonas reinhardtii. This screen led to identification of a gene encoding a protein designated Rubisco methyltransferase 2 (RMT2). Although homologous to methyltransferases, RMT2 has not been experimentally demonstrated to have methyltransferase activity. Furthermore, the rmt2 mutant was not compromised for Rubisco (first enzyme of Calvin-Benson Cycle) levels but did exhibit a marked decrease in accumulation/activity of photosystem I (PSI), which causes light sensitivity, with much less of an impact on other photosynthetic complexes. This mutant also shows increased accumulation of Ycf3 and Ycf4, proteins critical for PSI assembly. Rescue of the mutant phenotype with a wild-type (WT) copy of RMT2 fused to the mNeonGreen fluorophore indicates that the protein localizes to the chloroplast and appears to be enriched in/around the pyrenoid, an intrachloroplast compartment present in many algae that is packed with Rubisco and potentially hypoxic. These results indicate that RMT2 serves an important role in PSI biogenesis which, although still speculative, may be enriched around or within the pyrenoid. Significance StatementA high-throughput genetic screen was used to identify O2 sensitive mutants of Chlamydomonas reinhardtii (Chlamydomonas throughout) that experience elevated oxidative stress in the light relative to WT cells. Identification of genes altered in these mutants offers opportunities to discover activities that a) protect photosynthetic cells from oxidative damage, b) participate in rapid assembly of photosynthetic complexes, which would limit accessibility of intermediates to O2, and/or c) facilitate repair of damaged cellular complexes. A mutant from this screen disrupted for RMT2, originally described as encoding a Rubisco methyltransferase, was defective for PSI biogenesis. Additionally, RMT2 appears to be enriched in/around the pyrenoid, a chloroplast localized compartment harboring much of the Chlamydomonas Rubisco, raising the possibility that this compartment plays a role in PSI biogenesis.

plant biology↗

bioRxiv2023

Draft genome sequence of Microbacterium fakhimi sp. nov., a novel bacterium associated with the alga Chlamydomonas reinhardtii

Microbacterium fakhimi sp. nov. has been isolated from a contaminated algal culture (Chlamydomonas reinhardtii). Its genome has been fully sequenced (3,753,259 base pairs) and a tentative annotation is provided (3,704 genes). Both, genome information and growth tests suggest that M. fakhimi sp. nov. is auxotroph for biotin and thiamine and unable to use sulfate as sulfur (S) source. S-reduced forms, such as methionine and cysteine can support M. fakhimi sp. nov. growth. The potential biotechnological interest of this bacteria is discussed here and in a related research paper (Fakhimi et al., 2023b).

microbiology↗

bioRxiv2023

Draft genome sequence of Stenotrophomonas goyi sp. nov., a novel bacterium associated with the alga Chlamydomonas reinhardtii

Stenotrophomonas goyi sp. nov. has been isolated from a contaminated algal culture (Chlamydomonas reinhardtii). Its genome has been fully sequenced (4,487,389 base pairs) and a tentative annotation is provided (4,147 genes). The genome information suggests that S. goyi sp. nov. is unable to use sulfate and nitrate as sulfur and nitrogen sources, respectively. Growth tests have confirmed the dependence of the sulfur-containing amino acids methionine and cysteine. The potential biotechnological interest of this bacteria is discussed here and in a related research paper (Fakhimi et al., 2023b).

microbiology↗

bioRxiv2023

Chlamydomonas reinhardtii and Microbacterium fakhimi sp. nov., a mutualistic association that favor sustainable hydrogen production.

A multispecies bacterial community including Microbacterium forte sp. nov., Stenotrophomonas goyi sp. nov., and Bacillus cereus greatly promoted sustained hydrogen production by the microalga Chlamydomonas reinhardtii when cocultivated in mannitol- and yeast extract-containing medium (up to 313 mL{middle dot}L-1). Alga viability was also largely prolonged in the cocultures (>45 days) without any nutrient supplementation. Among the bacterial community, Microbacterium forte sp. nov. was the main responsible for the hydrogen production improvement. Nonetheless, the use of the entire bacterial community allowed a better growth of the alga during hydrogen production. Chlamydomonas reinhardtii and Microbacterium forte sp. nov. established a mutualistic association, based on the release of ammonium and acetic acid from the bacterium, while the alga provided sulfur-containing metabolites and complemented the bacterial auxotrophy for biotin and thiamine. This study uncovers the potential of the Chlamydomonas-bacteria consortia for durable and stable H2 production while allowing the simultaneous production of biomass.

microbiology↗

bioRxiv2022

Chlamydomonas reinhardtii triose-phosphate/phosphate translocator3 (TPT3): A major chloroplast shunt for the export of fixed carbon and reductant

Modulation of export of photoassimilates from the chloroplast is essential for controlling the distribution of fixed carbon in the cell and maintaining optimum photosynthetic rates. In this study we identified chloroplast triose phosphate/phosphate translocators 2 and 3 (CreTPT2 and CreTPT3) in the green alga Chlamydomonas reinhardtii that exhibited similar substrate specificities but were differentially expressed over the diel cycle. We focused mostly on analyzing CreTPT3 because of its high level of expression and the severe phenotype exhibited by tpt3 relative to the tpt2 mutants. Null mutants for CreTPT3 had a pleiotropic phenotype that impacted growth, photosynthetic activities, metabolite profiles, carbon partitioning, and organelle-specific accumulation of H2O2. These analyses demonstrated that CreTPT3 is a dominant conduit on the chloroplast envelope for the transport of photoassimilate. In addition, CreTPT3 can serve as a safety valve that moves excess reductant out of the chloroplast and appears to be essential for preventing the cells from experiencing oxidative stress and accumulating of reactive oxygen species, even under low/moderate light intensities. Finally, our studies indicate subfunctionalization of the CreTPT transporters and suggest that there are differences in managing the export of photoassimilates from the chloroplasts of Chlamydomonas and vascular plants.

plant biology↗