Plant J. 2023 Apr 3. doi: 10.1111/tpj.16231. Online ahead of print.

ABSTRACT

Carbonic anhydrases (CAs) are ubiquitous enzymes that accelerate the reversible conversion of CO₂ to HCO₃ ^- . The Arabidopsis genome encodes members of the α, β, and γ-CA families, and it has been hypothesized that βCA activity has a role in photosynthesis. In this work, we tested this hypothesis by characterizing the two plastidial βCAs, βCA1, and βCA5. We conclusively established that both proteins are localized in the chloroplast stroma and that the loss of βCA5 induced the expression of βCA1, supporting the existence of regulatory mechanisms to control the expression of stromal βCAs. We also established that βCA1 and βCA5 have markedly different enzymatic kinetics and physiological relevance. Specifically, we found that βCA5 had a first-order rate constant ~10-fold lower than βCA1 and that the loss of βCA5 is detrimental to growth and could be rescued by high CO₂ . Furthermore, we established that, while a βCA1 mutation showed near wild-type growth and no significant impact on photosynthetic efficiency, the loss of βCA5 markedly disrupted photosynthetic efficiency and light harvesting capacity at ambient CO₂ . Therefore, we conclude that in autotrophic growth, the loss of the more highly expressed βCA1 does not compensate for the loss of a less active βCA5, which in turn is involved in growth and photosynthesis at ambient CO₂ levels. These results lend support to the hypothesis that, in Arabidopsis, βCAs have non-overlapping roles in photosynthesis and identify a critical activity of stromal βCA5 and a dispensable role for βCA1.

PMID:37010739 | DOI:10.1111/tpj.16231

Chemistry. 2023 Apr 3:e202300854. doi: 10.1002/chem.202300854. Online ahead of print.

ABSTRACT

Invited for the cover of this issue are Christo Z. Christov and co-workers at Michigan Technological University, University of Oxford, and Michigan State University. The image depicts the oxygen diffusion channel in class 7 histone demethylase (PHF8) and ethylene-forming enzyme (EFE) and changes in the enzymes' conformations upon binding. Read the full text of the article at 10.1002/chem.202300138.

PMID:37009811 | DOI:10.1002/chem.202300854

Exp Cell Res. 2023 Mar 31;427(1):113571. doi: 10.1016/j.yexcr.2023.113571. Online ahead of print.

ABSTRACT

Nuclear speckles are subcellular structures originally characterized by punctate immunofluorescence staining of the monoclonal antibody SC35, which recognizes an epitope on SRRM2 (serine/arginine repetitive matrix protein 2) and Sfrs2, a member of the SR (serine/arginine-rich) family of splicing factors. Galectin-3 co-localizes with SC35 in nuclear speckles, which represent one group of nuclear bodies that include the nucleolus, Cajal bodies and gems, paraspeckles, etc. Although they appear to have well-delineated physical boundaries, these nuclear bodies are not membrane-bound structures but represent macromolecular assemblies arising from a phenomenon called liquid-liquid phase separation. There has been much recent interest in liquid phase condensation as a newly recognized mechanism by which a cell can organize and compartmentalize subcellular structures with distinct composition. The punctate/speckled staining of galectin-3 with SC3 demonstrates their co-localization in a phase-separated body in vivo, under conditions endogenous to the cell. The purpose of the present review is to summarize the studies that document three key features of galectin-3 for its localization in liquid phase condensates: (a) an intrinsically disordered domain; (b) oligomer formation for multivalent binding; and (c) association with RNA and ribonucleoprotein complexes.

PMID:37003559 | DOI:10.1016/j.yexcr.2023.113571

Sci Data. 2023 Mar 28;10(1):173. doi: 10.1038/s41597-023-01984-9.

ABSTRACT

This dataset contains ligand conformations and docking scores for 1.4 billion molecules docked against 6 structural targets from SARS-CoV2, representing 5 unique proteins: MPro, NSP15, PLPro, RDRP, and the Spike protein. Docking was carried out using the AutoDock-GPU platform on the Summit supercomputer and Google Cloud. The docking procedure employed the Solis Wets search method to generate 20 independent ligand binding poses per compound. Each compound geometry was scored using the AutoDock free energy estimate, and rescored using RFScore v3 and DUD-E machine-learned rescoring models. Input protein structures are included, suitable for use by AutoDock-GPU and other docking programs. As the result of an exceptionally large docking campaign, this dataset represents a valuable resource for discovering trends across small molecule and protein binding sites, training AI models, and comparing to inhibitor compounds targeting SARS-CoV-2. The work also gives an example of how to organize and process data from ultra-large docking screens.

PMID:36977690 | PMC:PMC10044124 | DOI:10.1038/s41597-023-01984-9

CBE Life Sci Educ. 2023 Jun;22(2):ar23. doi: 10.1187/cbe.20-01-0003.

ABSTRACT

Pressure gradients serve as the key driving force for the bulk flow of fluids in biology (e.g., blood, air, phloem sap). However, students often struggle to understand the mechanism that causes these fluids to flow. To investigate student reasoning about bulk flow, we collected students' written responses to assessment items and interviewed students about their bulk flow ideas. From these data, we constructed a bulk flow pressure gradient reasoning framework that describes the different patterns in reasoning that students express about what causes fluids to flow and ordered those patterns into sequential levels from more informal ways of reasoning to more scientific, mechanistic ways of reasoning. We obtained validity evidence for this bulk flow pressure gradient reasoning framework by collecting and analyzing written responses from a national sample of undergraduate biology and allied health majors from 11 courses at five institutions. Instructors can use the bulk flow pressure gradient reasoning framework and assessment items to inform their instruction of this topic and formatively assess their students' progress toward more scientific, mechanistic ways of reasoning about this important physiological concept.

PMID:36972334 | DOI:10.1187/cbe.20-01-0003