SUMMARY Regulatory T (Treg) cells suppress inflammatory immune responses and autoimmunity caused by self-reactive T cells. The key Treg cell transcription factor Foxp3 is downregulated during inflammation to allow for the acquisition of effector T cell-like functions. Here, we demonstrate that stress signals elicited by proinflammatory cytokines and lipopolysaccharide lead to the degradation of Foxp3 through the action of the E3 ubiquitin ligase Stub1. Stub1 interacted with Foxp3 to promote its K48-linked polyubiquitination in an Hsp70-dependent manner. Knockdown of endogenous Stub1 or Hsp70 prevented Foxp3 degradation. Furthermore, the overexpression of Stub1 in Treg cells abrogated their ability to suppress inflammatory immune responses in vitro and in vivo, and conferred a T helper 1 (Th1) cell-like phenotype. Our results demonstrate the critical role of the stress-activated Stub1-Hsp70 complex in promoting Treg cell inactivation, thus providing a potential therapeutic target for the intervention against autoimmune disease, infection and cancer.

Aroma is an important part of quality in table grape, but the key aroma compounds and the aroma series of table grapes remains unknown. In this paper, we identified 67 aroma compounds in 20 table grape cultivars; 20 in pulp and 23 in skin were active compounds. C6 compounds were the basic background volatiles, but the aroma contents of pulp juice and skin depended mainly on the levels of esters and terpenes, respectively. Most obviously, ‘Kyoho’ grapevine series showed high contents of esters in pulp, while Muscat/floral cultivars showed abundant monoterpenes in skin. For the aroma series, table grapes were characterized mainly by herbaceous, floral, balsamic, sweet and fruity series. The simple and visualizable aroma profiles were established using aroma fingerprints based on the aromatic series. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) showed that the aroma profiles of pulp juice, skin and whole berries could be classified into 5, 3, and 5 groups, respectively. Combined with sensory evaluation, we could conclude that fatty and balsamic series were the preferred aromatic series, and the contents of their contributors (β-ionone and octanal) may be useful as indicators for the improvement of breeding and cultivation measures for table grapes.

Circular RNAs (circRNAs) are widely distributed and play essential roles in a series of developmental processes, although none have been identified or characterized in grapevines (Vitis vinifera). In this study, we characterized the function of grape circRNA and uncovered thousands of putative back-splicing sites by global transcriptome analysis. Our results indicated that several reported circRNA prediction algorithms should be used simultaneously to obtain comprehensive and reliable circRNA predictions in plants. Furthermore, the length of introns flanking grape circRNAs was closely related to exon circularization. Although the longer introns flanking grape circRNAs appeared to circularize more efficiently, a 20-to 50-nt region seemed large enough to drive grape circRNA biogenesis. In addition, the endogenous introns flanking circularized exon(s) in conjunction with reverse complementary sequences could support the accurate and efficient circularization of various exons in grape, which constitutes a new tool for exploring the functional consequences caused by circRNA expression. Finally, we identified 475 differentially expressed circRNAs in grape leaves under cold stress. Overexpression of Vv-circATS1, a circRNA derived from glycerol-3-P acyltransferase, improved cold tolerance in Arabidopsis (Arabidopsis thaliana), while the linear RNA derived from the same sequence cannot. These results indicate the functional difference between circRNA and linear RNA, and provide new insight into plant abiotic stress resistance.

Forkhead box P3 (FOXP3)-positive Treg cells are crucial for maintaining immune homeostasis. FOXP3 cooperates with its binding partners to elicit Treg cells' signature and function, but the molecular mechanisms underlying the modulation of the FOXP3 complex remain unclear. Here we report that Deleted in breast cancer 1 (DBC1) is a key subunit of the FOXP3 complex. We found that DBC1 interacts physically with FOXP3, and depletion of DBC1 attenuates FOXP3 degradation in inflammatory conditions. Treg cells from Dbc1-deficient mice were more resistant to inflammationmediated abrogation of Foxp3 expression and function and delayed the onset and severity of experimental autoimmune encephalomyelitis and colitis in mice. These findings establish a previously unidentified mechanism regulating FOXP3 stability during inflammation and reveal a pathway for potential therapeutic modulation and intervention in inflammatory diseases.+ Treg cells are actively engaged in the prevention of autoimmunity and the mitigation of aberrant or excessive immune responses (1-3). The transcription factor Forkhead box P3 (denoted "FOXP3" in humans, and "Foxp3" in mice) is a well-characterized marker of Treg cells, and its expression typically is considered a requisite for Tregcell differentiation and function (4, 5). FOXP3 deficiency leads to the scurfy phenotype in mice and to the immune dysregulation, polyendocrinopathy, and enteropathy, X-linked syndrome in humans (6). Moreover, Treg-cell function is impaired in several autoimmune and inflammatory diseases, including colitis, rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus (7). Thus, the manipulation of Treg-cell function might provide a practical approach to the treatment of autoimmune and inflammatory diseases.Despite the central role of FOXP3 in Treg cells, many questions remain regarding the molecular mechanisms by which FOXP3 regulates Treg-cell function. Foxp3 protein is expressed transiently in CD4 + CD25− effector T cells upon T-cell receptor (TCR) stimulation but does not generate T cells with suppressive activity (8, 9). It has become evident that Foxp3 alone is insufficient to reproduce completely the differentiation and functional characteristics of Treg cells (10-12). FOXP3 binds with its partners to form multiple positive and negative feedback loops to regulate Treg-cell function subtly (13). FOXP3 interacts with FOXP1 to form heterodimers that promote FOXP3-mediated repression of IL-2 production (14). FOXP3 also binds with several nuclear factors, such as GATA3 (11, 15), RORγt (16), Eos (17), and RUNX1 (18), to drive the Treg cells' genetic program. FOXP3 function also is regulated at the posttranslational level. FOXP3 has been shown to interact with the acetyltransferase Tat-interaction protein 60 kDa (TIP60) to promote FOXP3 acetylation, which is required for Treg cells' suppressive function (19). P300 also regulates (11); therefore, many questions remain regarding the differential modulation of this complex and its effect on Treg-cell differenti...

Short hairpin (sh)RNAs delivered by recombinant adeno-associated viruses (rAAVs) are valuable tools to study gene function in vivo and a promising gene therapy platform. Our data show that incorporation of shRNA transgenes into rAAV constructs reduces vector yield and produces a population of truncated and defective genomes. We demonstrate that sequences with hairpins or hairpin-like structures drive the generation of truncated AAV genomes through a polymerase redirection mechanism during viral genome replication. Our findings reveal the importance of genomic secondary structure when optimizing viral vector designs. We also discovered that shDNAs could be adapted to act as surrogate mutant inverted terminal repeats (mTRs), sequences that were previously thought to be required for functional self-complementary AAV vectors. The use of shDNAs as artificial mTRs opens the door to engineering a new generation of AAV vectors with improved potency, genetic stability, and safety for both preclinical studies and human gene therapy.

Background: GATA3 is regulated both transcriptionally and post-translationally. GATA3 is important for the function of FOXP3 ϩ Treg cells.

Background: FOXP3 is a key transcription factor for the development and function of Tregs. Results: PIM1-mediated phosphorylation of FOXP3 at serine 422 decreased its DNA binding activity. Conclusion: PIM1 negatively regulates FOXP3-mediated transcriptional regulation and the suppressive activity of Tregs. Significance: PIM1 is a newly identified negative regulator of the immunosuppressive activity of Tregs.

Light is a key environmental factor affecting anthocyanin biosynthesis. Our previous study demonstrated that "Lanshan Hexian" is a light-sensitive eggplant cultivar, but its regulatory mechanism is unknown. Here, delphinidin-3-[4-(cis-p-coumaroyl)-rhamnosyl-glucopyranoside]-5-glucopyranoside and delphinidin-3-[4-(trans-p-coumaroyl)-rhamnosyl-glucopyranoside]-5-glucopyranoside were identified as the main anthocyanin components in Lanshan Hexian by ultra-performance liquid chromatography-tandem mass spectrometry. Three time points of anthocyanin accumulation, including the start point (0 day), fastest point (5 days), and highest point (12 day), were investigated by using ribonucleic acid sequencing and iTRAQ technology. The corresponding correlation coefficients of differentially expressed genes, and differentially expressed proteins were 0.6936, 0.2332, and 0.6672. Anthocyanin biosynthesis was a significantly enriched pathway, and CHI, F3H, 3GT, 5GT, and HY5 were regulated at both transcriptional and translational levels. Moreover, some transcription factors and photoreceptors may participate in light-induced anthocyanin biosynthesis like the known transcription factors MYB113 and TT8. The transient expression assay indicated that SmMYB35, SmMYB44, and a SmMYB86 isoform might involve in the light-induced anthocyanin biosynthesis pathway. Finally, a regulatory model for light-induced anthocyanin biosynthesis in eggplant was constructed. Our work provides a new direction for the study of the molecular mechanisms of light-induced anthocyanin biosynthesis in eggplant.