Elements In Promoters Of Genes Research Articles

Investigating the regulatory function of the ANO1-AS2 on the ANO1 gene in infertile men with asthenozoospermia and terato-asthenozoospermia

Long non-coding RNAs (lncRNAs) have a particular expression in the testicular tissue and exhibit a regulatory function on the reproduction system. ANO1-AS2 (linc02584), as an lncRNA is located near the anoctamin1 (ANO1) gene. ANO1 is an important component of the transmembrane system exhibiting expression modifications in the idiopathic infertile men. Therefore, the present study was conducted to investigate the relationship between ANO1-AS2 and ANO1 gene expression with sperm motility and morphology in the patients with asthenozoospermia (AZ) and terato- asthenozoospermia (TAZ).The study population included 32 patients with AZ, 35 patients with TAZ, and 34 people with normozoospermia (NZ, control). The expression levels of ANO1 gene and ANO1-AS2 in the spermatozoa were measured by the quantitative real-time polymerase chain reaction (PCR). Docking analysis was performed to investigate the interactions of the ANO1 gene promoter and intermediate elements with ANO1-AS2.ANO1 gene expression was significantly (P < 0.05) downregulated in the patients however; ANO1-AS2 expression was significantly upregulated (P < 0.05). The subsequent analysis confirmed the inverse correlation between ANO1 and ANO1-AS2. ANO1 gene expression level was significantly positively correlated with sperm motility and morphology (P < 0.05). Moreover, ANO1-AS2 expression showed an inverse correlation with sperm motility and morphology (P < 0.05). Docking analysis confirmed that ANO1-AS2 could stably interact with ANO1 gene promoter.In conclusion, ANO1-AS2 is likely to downregulate the ANO1 gene by interacting with ANO1 gene promoter, which can influence the sperm motility and morphology.

Enzymatic Protein Biopolymers as a Tool to Synthetize Eukaryotic Messenger Ribonucleic Acid (mRNA) with Uses in Vaccination, Immunotherapy and Nanotechnology

Multi-subunit enzymes are protein biopolymers that are involved in many cellular processes. The enzyme that carries out the process of transcription of mRNAs is RNA polymerase II (RNAPII), which is a multi-subunit enzyme in eukaryotes. This protein biopolymer starts the transcription from specific sites and is positioned by transcription factors, which form a preinitiation complex (PIC) on gene promoters. To recognize and position the RNAPII and the transcription factors on the gene promoters are needed specific DNA sequences in the gene promoters, which are named promoter elements. Those gene promoter elements can vary and therefore several kinds of promoters exist, however, it appears that all promoters can use a similar pathway for PIC formation. Those pathways are discussed in this review. The in vitro transcribed mRNA can be used as vaccines to fight infectious diseases, e.g., in immunotherapy against cancer and in nanotechnology to deliver mRNA for a missing protein into the cell. We have outlined a procedure to produce an mRNA vaccine against the SARS-CoV-2 virus, which is the causing agent of the big pandemic, COVID-19, affecting human beings all over the world. The potential advantages of using eukaryotic RNAPII to synthetize large transcripts are outlined and discussed. In addition, we suggest a method to cap the mRNA at the 5′ terminus by using enzymes, which might be more effective than cap analogs. Finally, we suggest the construction of a future multi-talented RNAPII, which would be able to synthetize large mRNA and cap them in the test tube.

Bioinformatics analysis of safflower WD40 transcription factor family genes

The WD40 transcription factor family is a gene superfamily widely found in eukaryotes, which is closely related to plant growth and development regulation. It has been reported that the WD40 transcription factor was involved in the synthesis of anthocyanins, which is one of the vital components of safflower flavonoid compounds. In this study, 40 CtWD40 members in the safflower genome were identified though bioinformatics tools and gene expression analysis methods. According to the WD40 protein sequence and phylogenetic characteristics of Arabidopsis and other plants, the safflower CtWD40 family was classified into 7 subfamilies. Conservative motif analysis was used to reveal the specific conserved motifs and gene structures of each subfamily member, and there exist a certain degree of similarities in the conserved motifs and gene structure between the closely related family members. Subsequently, the search for cis-acting elements of gene promoters found CtWD40-specific promoter elements, revealing the metabolic pathways which may involve. Next, enrichment of function analysis was employed to analyze the functional categories and cellular localization of the CtWD40 protein. Furthermore, the interactions between CtWD40 proteins predicted its potential regulatory function. Finally, 19 members of the safflower CtWD40 subfamily were analyzed by qRT-PCR, the result showed the expression patterns of these members were different in diverse tissue and flowering period. This study provides a basis for the functional and expression research of the CtWD40 genes.

Maize E2F transcription factors. Expression, association to promoters of S-phase genes and interaction with the RBR1 protein in chromatin during seed germination

For seed germination, it is necessary to restart the cell cycle, a process regulated at multiple levels including transcriptional control, that is executed by the E2F family of transcription factors. We identified 12 genes of the E2F family in maize that are expressed differentially during the first 28 h post imbibition (HAI). E2Fa/b1;1 and E2Fc proteins were characterized as an activator and a putative repressor respectively, both forming heterodimers with DPb2 that bind differentially to consensus E2F response elements in promoters of E2F target genes. Transcripts of target genes for these transcription factors accumulate during germination; in dry seeds E2Fc protein is enriched in the target promoters and is replaced by E2Fa/b1;1 as germination advances. RBR1 is found in the same promoters in non-imbibed and 28 HAI seeds, when DNA replication has concluded, and transcription of the E2F targets should stop. During germination promoters of these target genes seem to be decorated with histone marks related to relaxed chromatin structure. Therefore, E2Fs appear to occupy their target genes in a context of open chromatin, with RBR1 fine tuning the progression between the phases.

Genome wide analysis and functional identification of MdKCS genes in apple

Apple fruit is covered by cuticle wax, which plays important roles protecting fruits from adverse environmental conditions. β-Ketoacyl-CoA synthase (KCS) is the key rate-limiting enzyme in plant wax synthesis. In this study, we identified 28 KCS gene family members from apple (Malus × domestica Borkh.) by homology analysis. Multi-sequence alignment and phylogenetic analyses revealed that the 28 MdKCS genes were divided into four subgroups, including KCS1-like, FAE1-like, FDH-like, and CER6. A chromosomal localization analysis revealed that 27 apple KCS genes were located on 11 chromosomes, while MdKCS28 was localized to the unassembled genomic scaffold. Most of the MdKCS proteins were hydrophilic proteins and they had similar secondary and tertiary structures. The prediction of cis-acting elements of the MdKCS gene promoters suggested that the MdKCS genes may be widely involved in hormone signaling and the stress response. Furthermore, the quantitative real-time polymerase chain reaction results showed that eight MdKCS genes were highly expressed in the apple pericarp, and were significantly induced by drought, abscisic acid (ABA), and NaCl treatments. We transformed the MdKCS21 gene into apple calli, and found the MdKCS21 overexpressing transgenic apple calli exhibited higher tolerance to ABA treatment. Finally, the MdKCS proteins were localized to the endoplasmic reticulum and vacuolar membrane by confocal laser microscopy. This study established a foundation to further analyze the function of KCS genes and provided candidate genes for molecular improvement of wax content in apple.

First Expressed TFome of Physic Nut (Jatropha curcas L.) After Salt Stimulus

Physic nut (Jatropha curcas L.), a small oleaginous tree spontaneously occurring in arid and semi-arid tropical regions, is a sustainable and renewable energy source for biodiesel. However, the J. curcas yield in such areas should consider soil salinity and its consequences. Transcription factor (TF) proteins recognize cis-regulatory elements in promoters of genes to be expressed. In the present work, differentially expressed genes (DEGs) encoding putative TFs from physic nut plants responding to NaCl (150 mM), after 3 h of exposition, covered 23 TF families. The expressed profiles of members from AP2/ERF and NAC families basically showed induction after the salt stimulus, while members of bHLH, FHY3-FAR1, and ARF families presented repression. Concerning the induced TF DEGs, the gene ontology (GO) enrichment analysis highlighted terms related to abiotic stress responses, while those terms representing the repressed TF DEGs stood out the basal metabolism. In turn, the TF enrichment analysis predicted those TFs targeting promoters of induced TF DEGs. Some of the enriched TFs may be good candidates as transgenes in transgenic events. Also, RT-qPCR analyses validated the up-regulation of six TF DEGs (RAV1, ERF9, ZAT12, PTI5, MYB340, and BZIP4) of eight candidates selected from the expressed TFome. The generated data could help breeders to better understand the molecular basis of physic nut plants responding to salinity, to select potential candidates for transgenic studies, as well as to develop functional molecular markers to assist selection steps in breeding programs.

The In Vitro Functional Impairment of Thyroid Hormone Receptor Alpha 1 Isoform Mutants Is Mainly Dictated by Reduced Ligand Sensitivity.

Background: Thyroid hormone (TH) acts on TH receptors (TRs) and regulates gene transcription by binding of TRs to TH response elements (TREs) in target gene promoters. The transcriptional activity of TRs is modulated by interactions with TR-coregulatory proteins. Mutations in TRα cause resistance to thyroid hormone alpha (RTHα). In this study, we analyzed if, beyond reduced triiodothyronine (T3) affinity, altered interactions with cofactors or different TREs could account for the differential impaired transcriptional activity of different mutants. Methods: We evaluated four mutants derived from patients (D211G, M256T, A263S, and R384H) and three artificial mutants at equivalent positions in patients with RTHβ (T223A, L287V, and P398H). The in vitro transcriptional activity was evaluated on TRE-luciferase reporters (DR4, IR0, and ER6). The affinity for T3 and interaction with coregulatory proteins (nuclear receptor corepressor 1 [NCoR1] and steroid receptor coactivator 1 [SRC1]) were also determined. Results: We found that the affinity for T3 was significantly reduced for all mutants, except for TRα1-T223A. The reduction in the T3 sensitivity of the transcriptional activity on three TREs, the dissociation of the corepressor NCoR1, and the association of the coactivator SRC1 recruitment for each mutant correlated with the reduced affinity for T3. We did not observe mutation-specific alterations in interactions with cofactors or TREs. Conclusions: In summary, the degree of impaired transcriptional activity of mutants is mainly determined by their reduced affinity for T3.

Diagnostic kit for rice blight resistance

Blight-resistant rice lines are the most effective solution for bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo). Key resistance mechanisms involve SWEET genes as susceptibility factors. Bacterial transcription activator-like (TAL) effectors bind to effector-binding elements (EBEs) in SWEET gene promoters and induce SWEET genes. EBE variants that cannot be recognized by TAL effectors abrogate induction, causing resistance. Here we describe a diagnostic kit to enable analysis of bacterial blight in the field and identification of suitable resistant lines. Specifically, we include a SWEET promoter database, RT–PCR primers for detecting SWEET induction, engineered reporter rice lines to visualize SWEET protein accumulation and knock-out rice lines to identify virulence mechanisms in bacterial isolates. We also developed CRISPR–Cas9 genome-edited Kitaake rice to evaluate the efficacy of EBE mutations in resistance, software to predict the optimal resistance gene set for a specific geographic region, and two resistant ‘mega’ rice lines that will empower farmers to plant lines that are most likely to resist rice blight.

Genome-Wide Identification and Characterization of the Vacuolar H+-ATPase Subunit H Gene Family in Crop Plants.

The vacuolar H+-ATPase (V-ATPase) plays many important roles in cell growth and in response to stresses in plants. The V-ATPase subunit H (VHA-H) is required to form a stable and active V-ATPase. Genome-wide analyses of VHA-H genes in crops contribute significantly to a systematic understanding of their functions. A total of 22 VHA-H genes were identified from 11 plants representing major crops including cotton, rice, millet, sorghum, rapeseed, maize, wheat, soybean, barley, potato, and beet. All of these VHA-H genes shared exon-intron structures similar to those of Arabidopsis thaliana. The C-terminal domain of VHA-H was shorter and more conserved than the N-terminal domain. The VHA-H gene was effectively used as a genetic marker to infer the phylogenetic relationships among plants, which were congruent with currently accepted taxonomic groupings. The VHA-H genes from six species of crops (Gossypium raimondii, Brassica napus, Glycine max, Solanum tuberosum, Triticum aestivum, and Zea mays) showed high gene structural diversity. This resulted from the gains and losses of introns. Seven VHA-H genes in six species of crops (Gossypium raimondii, Hordeum vulgare, Solanum tuberosum, Setaria italica, Triticum aestivum, and Zea mays) contained multiple transcript isoforms arising from alternative splicing. The study of cis-acting elements of gene promoters and RNA-seq gene expression patterns confirms the role of VHA-H genes as eco-enzymes. The gene structural diversity and proteomic diversity of VHA-H genes in our crop sampling facilitate understanding of their functional diversity, including stress responses and traits important for crop improvement.

Ethanol promotes differentiation of embryonic stem cells through retinoic acid receptor-γ

Ethanol (EtOH) is a teratogen, but its teratogenic mechanisms are not fully understood. The alcohol form of vitamin A (retinol/ROL) can be oxidized to all-trans-retinoic acid (RA), which plays a critical role in stem cell differentiation and development. Using an embryonic stem cell (ESC) model to analyze EtOH's effects on differentiation, we show here that EtOH and acetaldehyde, but not acetate, increase differentiation-associated mRNA levels, and that EtOH decreases pluripotency-related mRNAs. Using reporter assays, ChIP assays, and retinoic acid receptor-γ (RARγ) knockout ESC lines generated by CRISPR/Cas9 and homologous recombination, we demonstrate that EtOH signals via RARγ binding to RA response elements (RAREs) in differentiation-associated gene promoters or enhancers. We also report that EtOH-mediated increases in homeobox A1 (Hoxa1) and cytochrome P450 family 26 subfamily A member 1 (Cyp26a1) transcripts, direct RA target genes, require the expression of the RA-synthesizing enzyme, aldehyde dehydrogenase 1 family member A2 (Aldh1a2), suggesting that EtOH-mediated induction of Hoxa1 and Cyp26a1 requires ROL from the serum. As shown with CRISPR/Cas9 knockout lines, the retinol dehydrogenase gene Rdh10 and a functional RARE in the ROL transporter stimulated by retinoic acid 6 (Stra6) gene are required for EtOH induction of Hoxa1 and Cyp26a1 We conclude that EtOH stimulates stem cell differentiation by increasing the influx and metabolism of ROL for downstream RARγ-dependent transcription. In stem cells, EtOH may shift cell fate decisions to alter developmental outcomes by increasing endogenous ROL/RA signaling via increased Stra6 expression and ROL oxidation.

Bioinformatics Analysis Makes Revelation to Potential Properties on Regulation and Functions of Human Sox2.

Sex determining region Y-box2 (Sox2) is a transcription factor that is essential for maintaining self-renewal or pluripotency of undifferentiated embryonic stem cells. The expression and distribution of Sox2 in tumor tissues have been extensively recorded, which are related to the progression and metastasis of tumor. However, a complete mechanistic understanding of Sox2 regulation and function remains to be studied. Herein, we show new potential properties of Sox2 regulation and functions from bioinformatics analysis. We use numerous algorithms to characterize the Sox2 gene promoter elements and the Sox2 protein structure, physio-chemical, localization properties and its evolutionary relationships. The expression of Sox2 is regulated by a diverse set of transcription factors and associated with the levels of methylation of CpG Islands in promoters. The structural properties of Sox2 indicate that Sox2 expresses as a stem cell marker in a variety of stem cells. Sox2 together with other transcription factors or proteins regulate the expression of downstream target genes, which makes a great difference to the biological function of stem cells. Not only stem cells, Sox2 also play an important role in tumor cells. In conclusion, this information from bioinformatics analysis will help to understand Sox2 regulation and functions better in future attempts.

TALE-carrying bacterial pathogens trap host nuclear import receptors for facilitation of infection of rice.

SummaryMany plant‐pathogenic Xanthomonas rely on the secretion of virulence transcription activator‐like effector (TALE) proteins into plant cells to activate plant susceptibility genes to cause disease. The process is dependent on the binding of TALEs to specific elements of host target gene promoters in the plant nucleus. However, it is unclear how TALEs, after injection into host cells, are transferred from the plant cytoplasm into the plant nucleus, which is the key step of successful pathogen infection. Here, we show that the host plant cytoplasm/nuclear shuttle proteins OsImpα1a and OsImpα1b are key components for infection by the TALE‐carrying bacterial pathogens Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xoc), the causal agents of bacterial leaf blight and bacterial leaf streak, respectively, in rice. Direct interaction between the second nuclear localization signal of TALEs of Xoo or Xoc and OsImpα1a or OsImpα1b is required for the transportation of TALEs into the nucleus. Conversely, suppression of the expression of OsImpα1a and OsImpα1b genes attenuates the shuttling of TALEs from the cytoplasm into the nucleus and the induction of susceptibility genes, thus improving the broad‐spectrum disease resistance of rice to Xoo and Xoc. These results provide an applicable strategy for the improvement of resistance to TALE‐carrying pathogens in rice by moderate suppression of the expression of plant nuclear import receptor proteins.

Analysing structural diversity of seed storage protein gene promoters: Buckwheat a case study

Multiple sequence alignment of 5’UTR of SSP genes from accessions of Fagopyrum esculentumrevealed the invariant nature of sequences with the transcription start site at P761and TATA box located -30bp upstream the TSS. Other cis-elements identified in the sequences included the legumin box (-581, -524, -184, -135, -91), the -131 prolamin box, DOF element (-718, -649, -540,-432, -272,-225, -128) and CAAT box (-692, -530, -475, -411, -282, -168, -54). Other elements identified included those involved in abscisic acid signallingviz., ABI3 at P-470,-95,-68,RAV1 at P-694and -543and AGL15 at P-671. A comparative analysis of regulatory elements of SSP gene promoters of distantly related species the presence of five cis-regulatory elements viz. TATA BOX, E-BOX, RY- element, CAAT box and the Endosperm box, which interplay in seed specific SSP gene expression. Other modulators influencing seed specific gene expression detected in the sequences included the ABA-responsive elements ABI3, RAV1 and AGL15 which play an integral role in seed maturation. Identification of potential nucleosome binding sites in SSP gene promoters of Cicer arietinum, Brassica napus, B. campestris, Vicia faba, and Pisum sativumat positions 78, 635, 195, 112 and 152 respectively surmises the spatial fine tuning of SSP gene transcriptional regulation in these species. On the other hand, absence of nucleosome binding sites in the promoters of Fagopyrum esculentum, Zea mays, Avena sativa, Triticum aestivum and Oryza sativamay indicate relatively easier access of transcription factors to the proximal promoter, thereby providing higher level of gene expression.

Analysis of cysteine protease inhibitor gene (BmCPI) promoter activity in silkworms using bac-to-bac baculovirus systems.

A cysteine protease inhibitor of Bombyx mori (BmCPI) plays an important role in pupation, molting, and dissociation of tissues. The present study identified and analyzed the BmCPI promoter region to better understand its functional regulatory mechanisms. Eight promoter fragments of different lengths were analyzed using an improved Bac-to-Bac expression system. Luciferase activities were investigated both in BmE cells and larval organisms after infection with the Bac-to-Bac system, and similar changes in activity were observed in both models. Strong activity was detected in the longest promoter (2005 bp, -1969 to +36), and activity changed significantly with truncation of promoter length. An electrophoretic mobility shift assay showed that the promoter region from -32 to +6 bp played a critical role in activating the downstream gene promoter element, where some potential elements were also predicted by informatics tools. The findings offer a basic reference for the mechanism of transcriptional regulation of BmCPI.

Genome-Wide Identification, Characterization and Expression Analyses of Heat Shock Protein-Related Genes in a Highly Invasive Ascidian Ciona savignyi.

Biological response to rapid changing environments is an outstanding research question in ecology and evolution. Biological invasions provide excellent “natural” experiments to study such a complex response process, as invaders often encounter rapidly changing environments during biological invasions. The regulation of heat shock proteins (Hsp) is a common pathway responsible for various environmental stresses; however, the comprehensive study on Hsp system across the whole genome and potential roles in determining invasion success are still largely unexplored. Here, we used a marine invasive model ascidian, Ciona savignyi, to investigate transcriptional response of Hsp-related genes to harsh environments. We identified 32 genes, including three Hsp20, six Hsp40, ten Hsp60, eight Hsp70, three Hsp90, one Hsp100, and one heat shock transcription factor (Hsf), across the whole genome of C. savignyi. We further characterized gene structure and protein motifs, and identified potential heat shock elements (HSEs) in promoters of Hsp genes. The expression analysis showed that most Hsp genes, but not all, were involved in transcriptional response to temperature and salinity challenges in a duration- and stress-specific pattern, and the maximum amplitude of induction occurred in Hsp70-4 after 1-h of high temperature treatment. However, the Hsf gene was scarcely induced and limited interactions were predicted between Hsp and Hsf genes. Our study provide the first systematic genome-wide analysis of Hsp and Hsf family in the marine invasive model ascidian, and our results are expected to dissect Hsp-based molecular mechanisms responsible for extreme environmental adaptation using Ciona as a model system.

PO-260 Anti-lymphoma activity of novel selective glucocorticoid receptor agonists (SEGRAs) in vitro and in vivo

IntroductionGlucocorticoids (GCs) are widely used in blood cancer treatment; although, they cause metabolic disorders. Biological response to GCs is mediated by glucocorticoid receptor (GR) regulating gene expression via transactivation (TA), which requires GR binding to GC-responsive elements in gene promoters, and transrepression (TR), negative interaction between GR and transcription factors. TR mediates anticancer effects of GR, while side effects are associated with GR TA. Selective GR agonists (SEGRAs) that preferentially activate GR TR could be a better option for cancer treatment. One of well characterised SEGRAs is 2-(4-acetoxyphenyl)−2-chloro-N-methylethylammonium-chloride, or CpdA, isolated from Namibian shrub Salsola tuberculatiformis. CpdA demonstrated anticancer activity in vitro and in vivo. We extended SEGRA list by synthesis of CpdA enantiomers and its chemical derivatives.Material and methodsSynthesis of (S) and (R)-CpdA was based on Sharpless asymmetric dihydroxylation. Chemical analogues of CpdA, CpdA01-08, were designed by appending of bulky substituent into benzene ring and to nitrogen atom or alkylation of carbon atom adjacent to chlorine atom. All experiments in vitro were carried out on Granta (lymphoma) and CEM (leukaemia) cells. Cells were treated with Dex, CpdA, (R) and (S)-CpdA, CpdA01-08. Effects on cell growth were evaluated by cell counting and flow cytometry. Affinity to GR was measured by competitive binding assay. Gene expression was measured by qPCR and Western blotting. GR and NF-kB activity was evaluated using Luciferase reporter analysis. Anticancer effect in vivo was determined using the model of murine lymphoma P388.Results and discussionsThe most cytotoxic compounds among 10 newly synthesised, CpdA03 and CpdA05, demonstrated the highest affinity to GR. They induced GR TR but not TA and proved their SEGRA properties. Effect of CpdA enantiomers on cell growth and survival was not significantly different from Dex and CpdA. CpdA03, cytotoxic SEGRA with the highest affinity to GR, comparable with DEX and CpdA, was tested in vivo for evaluation its anti-lymphoma activity, and demonstrated 3-fold decrease of tumour size in comparison with 2–2,5-fold decrease after Dex or CpdA treatment.ConclusionThe design of synthesis and evaluation of anticancer properties of new SEGRA are provided. According to our data one novel SEGRA, CpdA03, is perspective for further investigation as anti-lymphoma drug with reduced side effects.Study is funded by Russian Science Foundation No. 17-75-20124.

PO-173 Sialylation of integrin alpha 2 promotes ovarian cancer cells metastasis

IntroductionThe emergence of migratory cell populations within solid tumours is a critical early stage during cancer metastasis. We have previously reported the enhanced migratory properties of one such population, marked by CD66 expression, in cervical cancers. It is not clear what mechanisms drive such migratory populations, and how these cells would retain a memory of initial migration-inducing cues during metastasis. Here, we sought to assess whether an altered heterochromatin state, controlled by chromatin regulator Suv39H1, drives migratory populations in cervical cancer.Material and methodsTo avoid an assumption regarding the role of an Epithelial to Mesenchymal Transition (EMT) in migration, cervical cancer cell line SiHa was sorted based on migratory ability in vitro. Following this, Suv39H1 function was assessed in vitro using RNAi. These approaches were complemented by a clinicopathological approach, using histopathology of clinical biopsies to examine cancer progression, along with a meta-analysis of RNA-Seq and clinical data obtained from The Cancer Genome Atlas (TCGA). Lastly, to infer putative mechanisms involved, RNA-Seq, confocal imaging and H3K9me3 ChIP-Seq were performed on sorted migrated cell populations.Results and discussionsWe observe that migrated populations show low Suv39H1, and that Suv39H1 knockdown enhances cell migration. Using histopathology, we observed the proliferative expansion of a Suv39H1high pool of cells in early stages of clinical progression, followed by the emergence of a sarcomatoid, migratory Suv39H1low cell population in advanced stages. Additionally, a meta-analysis of TCGA data revealed that low tumoural Suv39H1 also correlated with lower patient survival, and with gene expression signatures of migration, TGF-β signalling, and links with a CD66 cell signature. Lastly, using genome wide profiling and imaging on migrated populations in vitro, we observed that these populations show Suv39H1-linked migratory expression signatures, decondensed nuclei and a broad loss of H3K9me3, particularly across gene promoter elements. Our findings suggest a role for broad Suv39H1-mediated heterochromatin changes in regulating fate plasticity in migrated populations, contrasting with locus-specific heterochromatin changes previously described in EMT-based models.ConclusionThus, we report a Suv39H1-low heterochromatin state as a driver of migratory cell populations in cervical cancers. The understanding of such drivers may be valuable in efforts to develop anti-metastatic strategies.

Bdp1 interacts with SNAPc bound to a U6, but not U1, snRNA gene promoter element to establish a stable protein-DNA complex.

In metazoans, U6 small nuclear RNA (snRNA) gene promoters utilize a proximal sequence element (PSE) recognized by the small nuclear RNA-activating protein complex (SNAPc). SNAPc interacts with the transcription factor TFIIIB, which consists of the subunits TBP, Brf1 (Brf2 in vertebrates), and Bdp1. Here, we show that, in Drosophila melanogaster, DmSNAPc directly recruits Bdp1 to the U6 promoter, and we identify an 87-residue region of Bdp1 involved in this interaction. Importantly, Bdp1 recruitment requires that DmSNAPc be bound to a U6 PSE rather than a U1 PSE. This is consistent with the concept that DmSNAPc adopts different conformations on U6 and U1 PSEs, which lead to the subsequent recruitment of distinct general transcription factors and RNA polymerases for U6 and U1 gene transcription.

Co-expression network analysis and cis-regulatory element enrichment determine putative functions and regulatory mechanisms of grapevine ATL E3 ubiquitin ligases

Arabidopsis thaliana Toxicos en Levadura (ATL) proteins are a subclass of the RING-H2 zinc finger binding E3 ubiquitin ligases. The grapevine (Vitis vinifera) ATL family was recently characterized, revealing 96 members that are likely to be involved in several physiological processes through protein ubiquitination. However, the final targets and biological functions of most ATL E3 ligases are still unknown. We analyzed the co-expression networks among grapevine ATL genes across a set of transcriptomic data related to defense and abiotic stress, combined with a condition-independent dataset. This revealed strong correlations between ATL proteins and diverse signal transduction components and transcriptional regulators, in particular those involved in immunity. An enrichment analysis of cis-regulatory elements in ATL gene promoters and related co-expressed genes highlighted the importance of hormones in the regulation of ATL gene expression. Our work identified several ATL proteins as candidates for further studies aiming to decipher specific grapevine resistance mechanisms activated in response to pathogens.

Identification of genetic elements in metabolism by high-throughput mouse phenotyping

Metabolic diseases are a worldwide problem but the underlying genetic factors and their relevance to metabolic disease remain incompletely understood. Genome-wide research is needed to characterize so-far unannotated mammalian metabolic genes. Here, we generate and analyze metabolic phenotypic data of 2016 knockout mouse strains under the aegis of the International Mouse Phenotyping Consortium (IMPC) and find 974 gene knockouts with strong metabolic phenotypes. 429 of those had no previous link to metabolism and 51 genes remain functionally completely unannotated. We compared human orthologues of these uncharacterized genes in five GWAS consortia and indeed 23 candidate genes are associated with metabolic disease. We further identify common regulatory elements in promoters of candidate genes. As each regulatory element is composed of several transcription factor binding sites, our data reveal an extensive metabolic phenotype-associated network of co-regulated genes. Our systematic mouse phenotype analysis thus paves the way for full functional annotation of the genome.