PMC

Differences between promoter activity and clonal sectors in leaves. a–c Dynamic expression pattern of the DRNL::GUS transgene as indicated on each picture for young leaf stages. d Sector distribution in seedlings expressing driver/reporter combinations sprayed with DEX when the cotyledons were fully opened. GUS sectors were absent from the two first leaves initiated in the embryo, and according to the biological half-life and threshold levels of DEX, DEX-dependent sectors were restricted to the next five successive leaves. Numerous sectors were observed in leaves 4–6, of which a substantial fraction (in purple) exhibited asymmetric apical domain sectors. All subsequent panels depict DEX-dependent clonal sectors due to DRNL::CRE-GR activity in driver/reporter combinations. e Two successive young leaves from the same seedling with representative clonal sectors in young leaf primordia. Note that in contrast to the DRNL::GUS pattern in a or b, the lateral lamina domains are free of GUS staining, whereas staining that includes the mid-rib in the younger or the apical sector and hydathodes in the older leaflet coincide with DRNL promoter activity. f–k A selection of asymmetric sectors and independent cellular trajectories in lamina halves. f Half-lamina early sector along the mid-rib (left) and an independent apical sector (right); g mid-rib sector with differential expansion in the apical domain; h differently shaped sectors on either side of the mid-rib; note the absence of expression in a small central region to the left of the mid-rib and the sector at the leaf margin to the right. i Split mid-rib sector and discrete GUS-stained cellular trajectories to either side of the mid-rib below a symmetrical wedge-shaped apical sector. j An asymmetric apical sector that in contrast to sectors in i or k includes the epidermal layer and hydathode sectors at either margin. k A split mid-rib sector that centrally extends into the left lamina half; note the GUS staining in secondary veins in the apical domain. l–o Less-frequent sectors observed in basal lamina domains, which potentially relate to transient DRNL promoter activity during early leaf development (compare with DRNL::GUS patterns in a–c. p Close-up of a hydathode sector that includes the terminal part of the connected vein. q Sector that includes a leaf serration. r–u Hydathode-associated sectors with increasing leaf age from r to u at the time of DEX application; note the absence of an apical sector and small sector size (u). v Two distinct clonal sectors associated with a single hydathode suggest independent cellular events

Identification of an allosteric sector in Hsp70 proteins. (A) A histogram of the Hsp70/110 sequences projected on a top axis of sequence variation derived from singular value decomposition and independent component analysis (see Materials and methods). This axis separates family into two sub-families that correspond to the allosteric Hsp70s and the non-allosteric Hsp110s. (B) A histogram of Hsp70 residues projected on the corresponding axis of positional co-evolution, showing that a small fraction of residues is largely responsible for the sequence divergence shown in panel A (115 sector out of 605 total residues or ∼20%). This defines the Hsp70 sector. The sector nearly equally comprises residues from the nucleotide-binding domain (blue, 56 residues) and the substrate-binding domain (green, 59 residues). (C) The Hsp70 sector mapped onto a model of the ATP-bound state of E. coli DnaK. The sector forms a physically contiguous group of residues that connect the ATP-binding site in the nucleotide-binding domain (pale blue) to the substrate-binding pocket of the substrate-binding domain (yellow) through the binding interface between the two domains. Sector positions are represented as spheres and colored as in panel B. Source data is available for this figure at www.nature.com/msb.

Growth rate on the number of admissions for NHS-funded care for NHS hospitals and independent sector providers (ISPs) by sustainability and transformation partnerships (STP)—April–July 2019 vs April-–July 2020 (%). NHS, National Health Service.

A fraction of Gcn4p target genes is induced by 3AT in gcn4Δ cells. (A) Venn diagram depicting the overlap among 613 genes for which statistically significant data were obtained (P ≤ 0.05) and that showed an induction ratio of ≥2 in the WT ± 3AT (set C), GCN4/gcn4Δ, or gcn4Δ ± 3AT experiments. There are 229 Gcn4p target genes (sectors A and B) found at the intersection between 462 genes induced by 3AT in the WT (sectors A, B, C, and D) and 311 genes showing Gcn4p dependence for induction in the GCN4/gcn4Δ experiment (sectors F, B, and A). There were 280 genes induced by 3AT in the gcn4Δ ± 3AT experiment (sectors A, C, and E). The 151 genes in sector B are Gcn4p targets that were not induced by 3AT in the gcn4Δ strain and thus are completely dependent on Gcn4p for induction. Sector A contains the subset of Gcn4p target genes that were also induced by 3AT in the gcn4Δ mutant. These 78 genes are subject to dual regulation and require Gcn4p only for maximal induction by 3AT. Sector C is comprised of the 133 genes induced by 3AT in both gcn4Δ and WT cells which showed little or no dependence on Gcn4p in the GCN4/gcn4Δ experiment (induction ratio of <2-fold). These genes are induced by 3AT independently of Gcn4p. Thus, ∼50% of the genes induced by 3AT in the WT are designated Gcn4p targets because of their dependence on Gcn4p for maximal induction ([A + B]/[A + B + C + D]), and 34% of these target genes (A/[A + B]) can be induced by 3AT even in the absence of Gcn4p. The genes in sector F showed greater expression in WT cells than in gcn4Δ cells in the presence of 3AT but were not induced by 3AT in the WT. This class includes additional Gcn4p targets that depend on Gcn4p primarily to prevent repression in histidine-starved cells (see text). The genes in sector E were induced only in gcn4Δ cells and thus belong to the class of genes induced by 3AT independently of Gcn4p. The results obtained for the 100 genes in sector D marked with an asterisk are paradoxical in that any genes induced by 3AT in the WT should be either dependent on (sector A or B) or independent of (sector C) Gcn4p for their induction. Close examination of this group revealed that 28 genes were induced only in the single WT ± 3AT experiment used to construct this diagram (data set C) but not in other experiments of the same type (data set A, B, or D); hence, these 28 genes probably are not 3AT inducible. Fifty of the remaining genes in sector D showed induction ratios between 1.5 and 2.0 in the GCN4/gcn4Δ experiment and most likely belong to sector A or B, comprising the Gcn4p targets. Fifteen of the remaining genes had induction ratios between 1.5 and 2.0 in the gcn4Δ ± 3AT experiment and probably belong to sector C or E, comprising the Gcn4p-independent 3AT-inducible genes. Among the 133 genes assigned to sector C, 53 had induction ratios between 1.5- and 2-fold in the GCN4/gcn4Δ experiment, suggesting that they belong to sector A: Gcn4p targets that are also inducible by a Gcn4p-independent mechanism. (B) Color display plot of the expression ratios of selected Gcn4p target genes that were induced ≥1.9-fold by 3AT in the gcn4Δ strain. The log₁₀ ratios of expression are depicted using the color code described for Fig. , with the brightest red depicting log₁₀ ratios of ≥1.0 and black signifying no significant change in expression (P > 0.05). Data were taken from the different experiments listed across the top (as defined in Fig. and Table ) for the genes listed on the right.