Biology subjects

Vinyard, D. J.

Publications and source records attributed to Vinyard, D. J..

3 recordsLinked to original sources

bioRxiv2026

Cryo-EM structures of photosystem I with alternative quinones reveals new insight into cofactor selectivity

Quinones are an integral component of electron transfer processes in photosynthetic and mitochondrial respiratory proteins. One such photosynthetic protein, Photosystem I, is an essential photooxidoreductase found in all oxygenic phototrophs. To better understand quinone chemistry and to form a basis for protein engineering, the menB gene in the model cyanobacterium Synechocystis sp. PCC 6803 was interrupted, blocking the biosynthesis of phylloquinone and causing it to be replaced by exchangeable plastoquinone-9 in the A1A and A1B quinone-binding sites of Photosystem I. This genetic variant has been instrumental in bioenergy research, enabling incorporation of a range of substituted and isotopically labeled quinones. Despite numerous valuable studies, the interpretation of biophysical data has been limited by a lack of structural data. To address this, we present the high-resolution cryo-EM structures of Photosystem I from the {Delta}menB variant containing (a) exchangeable plastoquinone-9 and (b) exogenously added 2-ethyl-1,4-napthoquinone at 1.90- and 2.05-[A] resolution, respectively. Unexpectedly, the quinones in the A1A and A1B sites of Photosystem I, previously believed to have similar binding affinities, are found to be asymmetric in their ability to bind and exchange plastoquinone-9. This work reveals new and important insight into the molecular basis for Photosystem I activity in the {Delta}menB variant, the power of metabolic plasticity to maintain protein stability, and the requirement for protein instability to facilitate ligand exchange.

biochemistry↗

bioRxiv2024

Enhancing Photosynthesis under Salt Stress via Directed Evolution in Cyanobacteria

A key aspect of enhancing photosynthesis is improving its recycling kinetics, enabling swift resumption of photochemical quenching following environmental disruptions or stress. Salt stress exacerbates high light stress in cyanobacteria and leads to severe yield losses in crop plants. Genetic traits that confer salt tolerance without compromising photosynthetic performance are essential for improving photosynthesis under these conditions. Here we applied accelerated evolution in Synechococcus elongatus PCC 7942 by conditionally suppressing its methyl-directed mismatch repair system to obtain beneficial genetic traits for enhanced photosynthesis under salt stress. Screening over 10,000 mutants, we isolated eight strains with increased biomass or sucrose productivity under salt stress. Genome sequencing revealed an average of 8-20 single nucleotide polymorphisms (SNPs) or indels per genome. Notably, mutations in the photosystem II (PSII) reaction center D1 gene, resulting in the amino acid changes L353F, I358N, and H359N at the carboxyl terminus of the pre-D1 (pD1) protein, improve photosynthesis under salt and combined salt and light stress by potentially accelerating D1 maturation during PSII repair. Phylogenetic analysis of pD1 across cyanobacteria and red algae highlights the broad significance of these adaptive genetic traits, underscoring the importance of leveraging evolutionary insights to improve photosynthesis under stress or fluctuating environments.

plant biology↗

bioRxiv2024

Studies of CrHCF244 reveal similarities and differences in psbA translation between Chlamydomonas reinhardtii and Arabidopsis thaliana

Translation of psbA, the chloroplast gene that encodes the D1 subunit of Photosystem II (PSII), is important for both PSII biogenesis and repair. The translation of the psbA transcript in the chloroplast is under the control of nuclear gene products. Using a Chlamydomonas forward genetic screen and whole genome sequencing, we found a mutant defective in PSII activity and mapped the causative gene to be the homolog of Arabidopsis High Fluorescence (HCF244) gene, or CrHCF244. We then demonstrated that CrHCF244 is required for psbA translation in the alga, consistent with the function of HCF244 in Arabidopsis. The Arabidopsis HCF244 gene also partially complemented the algal mutant. These results experimentally support the functional conservation of the homologs in green algae and land plants. Intriguingly, the CrHCF244 mutant also exhibited a relatively high rate of suppressor mutants, pointing to the presence of alternative factor(s)/pathway(s) for D1 translation control. The establishment of CrHCF244 as a psbA translation factor in Chlamydomonas showed the similarities in psbA translation regulation in algae and plants. The future identification of the alternative factor(s) in this alga will provide insights on psbA translation in plants. HighlightWe identified CrHCF244 as a translation factor of psbA in Chlamydomonas. Arabidopsis HCF244 partially complements Chlamydomonas {Delta}CrHCF244 mutant, indicating semi-conservation of the function of this gene between organisms. Suppressor mutants of {Delta}CrHCF244 suggest the presence of alternative translation factors in psbA translation.

plant biology↗