The prefoldin complex (PFDc) was identified in humans as a co-chaperone of the cytosolic chaperonin T-COMPLEX PROTEIN RING COMPLEX (TRiC)/CHAPERONIN CONTAINING TCP-1 (CCT). PFDc is conserved in eukaryotes and is composed of subunits PFD1 to 6, and PFDc-TRiC/CCT folds actin and tubulins. PFDs also participate in a wide range of cellular processes, both in the cytoplasm and in the nucleus, and their malfunction causes developmental alterations and disease in animals and altered growth and environmental responses in yeast and plants. Genetic analyses in yeast indicate that not all their functions require the canonical complex. The lack of systematic genetic analyses in plants and animals, however, makes it difficult to discern whether PFDs participate in a process as the canonical complex or in alternative configurations, which is necessary to understand their mode of action. To tackle this question, and on the premise that the canonical complex cannot be formed if one subunit is missing, we generated an Arabidopsis (Arabidopsis thaliana) mutant deficient in the six prefoldins and compared various growth and environmental responses with those of the individual mutants. In this way, we demonstrate that the PFDc is required for seed germination, to delay flowering, or to respond to high salt stress or low temperature, whereas at least two PFDs redundantly attenuate the response to osmotic stress. A coexpression analysis of differentially expressed genes in the sextuple mutant identified several transcription factors, including ABA INSENSITIVE 5 (ABI5) and PHYTOCHROME INTERACTING FACTOR 4 (PIF4), acting downstream of PFDs. Furthermore, the transcriptomic analysis allowed assigning additional roles for PFDs, for instance, in response to higher temperature.
The cellular diversity and complexity of the kidney are on par with its physiological intricacy. Although our anatomical understanding of the different segments and their functions is supported by a plethora of research, the identification of distinct and rare cell populations and their markers remains elusive. Here, we leverage the large number of cells and nuclei profiles using single-cell (scRNA-seq) and single-nuclei (snRNA-seq) RNA-sequencing to build a comprehensive atlas of the adult mouse kidney. We created MKA (Mouse Kidney Atlas) by integrating 59 publicly available single-cell and single-nuclei transcriptomic datasets from eight independent studies. The atlas contains more than 140.000 cells and nuclei covering different single-cell technologies, age, and tissue sections. To harmonize annotations across datasets, we constructed a hierarchical model of the cell populations present in our atlas. Using this hierarchy, we trained a model to automatically identify cells in unannotated datasets and evaluated its performance against well-established methods and annotation references. Our learnt model is dynamic, allowing the incorporation of novel cell populations and refinement of known profiles as more datasets become available. Using MKA and the learned model of cellular hierarchies, we predicted previously missing cell annotations from several studies and characterized well-studied and rare cell populations. This allowed us to identify reproducible markers across studies for poorly understood cell types and transitional states.
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