Genome-Wide Identification and Expression Analysis of Hsf and Hsp Gene Families in Cucumber (Cucumis Sativus L.)

Heat shock proteins (Hsps) are molecular chaperones that participate in plants' response to environmental stresses, including heat stress, and also play an essential role in plant growth and development. Hsps expression is monitored and regulated by specific types of transcription factors known as heat shock factors (Hsfs). Although the role of Hsfs and Hsps in stress response has been investigated in some plants, their role is still poorly understood in cucumber (Cucumis sativus L.). To reveal the mechanisms of cucumber Hsf and Hsp coping with various stresses, the analyses of cucumber Hsf and Hsp gene families were conducted using bioinformatics-based methods. A total of 23 Hsfs and 72 Hsps were identified in the cucumber genome (v3.0), and the gene structure and motif composition are relatively conserved in each subfamily. At least 23 pairs of heat shock genes underwent gene duplication in cucumber. A cis-element analysis is implicit that CsHsfs and CsHsps possessed at least one hormone or stress response cis-element, suggesting that CsHsf and CsHsp genes could respond to different stress conditions. Heatmaps of the CsHsf and CsHsp gene families indicated that most heat shock genes were expressed in different tissues and organs. RNA-seq showed that most of the cucumber Hsf and Hsp genes are differentially expressed upon exposure to biotic and abiotic stresses. These results provide valuable information to clarify the evolutionary relationship between the CsHsf and CsHsp family and to facilitate the functional characterization of the CsHsf and CsHsp genes in future studies.

Cooperative Regulation of Campylobacter jejuni Heat-Shock Genes by HspR and HrcA

The heat-shock response is defined by the transient gene-expression program that leads to the rapid accumulation of heat-shock proteins. This evolutionary conserved response aims at the preservation of the intracellular environment and represents a crucial pathway during the establishment of host–pathogen interaction. In the food-borne pathogen Campylobacter jejuni two transcriptional repressors, named HspR and HrcA, are involved in the regulation of the major heat-shock genes. However, the molecular mechanism underpinning HspR and HrcA regulatory function has not been defined yet. In the present work, we assayed and mapped the HspR and HrcA interactions on heat-shock promoters by high-resolution DNase I footprintings, defining their regulatory circuit, which governs C. jejuni heat-shock response. We found that, while DNA-binding of HrcA covers a compact region enclosing a single inverted repeat similar to the so-called Controlling Inverted Repeat of Chaperone Expression (CIRCE) sequence, HspR interacts with multiple high- and low-affinity binding sites, which contain HspR Associated Inverted Repeat (HAIR)-like sequences. We also explored the DNA-binding properties of the two repressors competitively on their common targets and observed, for the first time, that HrcA and HspR can directly interact and their binding on co-regulated promoters occurs in a cooperative manner. This mutual cooperative mechanism of DNA binding could explain the synergic repressive effect of HspR and HrcA observed in vivo on co-regulated promoters. Peculiarities of the molecular mechanisms exerted by HspR and HrcA in C. jejuni are compared to the closely related bacterium H. pylori that uses homologues of the two regulators.

Transcriptome and translatome changes in germinated pollen under heat stress uncover roles of transporter genes involved in pollen tube growth

ABSTRACTPlant reproduction is one key biological process very sensitive to heat stress and, as a consequence, enhanced global warming poses serious threats to food security worldwide. In this work we have used a high-resolution ribosome profiling technology to study how heat affects both the transcriptome and the translatome of Arabidopsis thaliana pollen germinated in vitro. Overall, a high correlation between transcriptional and translational responses to high temperature was found, but specific regulations at the translational level were also present. We show that bona fide heat shock genes are induced by high temperature indicating that in vitro germinated pollen is a suitable system to understand the molecular basis of heat responses. Concurrently heat induced significant down-regulation of key membrane transporters required for pollen tube growth, thus uncovering heat-sensitive targets. We also found that a large subset of the heat-repressed transporters is specifically up-regulated, in a coordinated manner, with canonical heat-shock genes in pollen tubes grown in vitro and semi in vivo, based on published transcriptomes from Arabidopsis thaliana. Ribosome footprints were also detected in gene sequences annotated as non-coding, highlighting the potential for novel translatable genes and translational dynamics.

Salt stress in the renal tubules is linked to TAL-specific expression of uromodulin and an upregulation of heat shock genes

Previously, our comprehensive cardiovascular characterization study validated Uromodulin as a blood pressure gene. Uromodulin is a glycoprotein exclusively synthesized at the thick ascending limb of the loop of Henle and is encoded by the Umod gene. Umod−/− mice have significantly lower blood pressure than Umod+/+ mice, are resistant to salt-induced changes in blood pressure, and show a leftward shift in pressure-natriuresis curves reflecting changes of sodium reabsorption. Salt stress triggers transcription factors and genes that alter renal sodium reabsorption. To date there are no studies on renal transcriptome responses to salt stress. Here we aimed use RNA-Seq to delineate salt stress pathways in tubules isolated from Umod+/+ mice (a model of sodium retention) and Umod−/− mice (a model of sodium depletion) ± 300 mosmol sodium chloride ( n = 3 per group). In response to salt stress, the tubules of Umod+/+ mice displayed an upregulation of heat shock transcripts. The greatest changes occurred in the expression of: Hspa1a (Log2 fold change 4.35, P = 2.48 e−12) and Hspa1b (Log2 fold change 4.05, P = 2.48 e−12). This response was absent in tubules of Umod−/− mice. Interestingly, seven of the genes discordantly expressed in the Umod−/− tubules were electrolyte transporters. Our results are the first to show that salt stress in renal tubules alters the transcriptome, increasing the expression of heat shock genes. This direction of effect in Umod+/+ tubules suggest the difference is due to the presence of Umod facilitating greater sodium entry into the tubule cell reflecting a specific response to salt stress.