Reciprocal inhibition between TP63 and STAT1 regulates anti-tumor immune response through interferon-γ signaling in squamous cancer.

Squamous cell carcinomas (SCCs) are common and aggressive malignancies. Immune check point blockade (ICB) therapy using PD-1/PD-L1 antibodies has been approved in several types of advanced SCCs. However, low response rate and treatment resistance are common. Improving the efficacy of ICB therapy requires better understanding of the mechanism of immune evasion. Here, we identify that the SCC-master transcription factor TP63 suppresses interferon-γ (IFNγ) signaling. TP63 inhibition leads to increased CD8+ T cell infiltration and heighten tumor killing in in vivo syngeneic mouse model and ex vivo co-culture system, respectively. Moreover, expression of TP63 is negatively correlated with CD8+ T cell infiltration and activation in patients with SCC. Silencing of TP63 enhances the anti-tumor efficacy of PD-1 blockade by promoting CD8+ T cell infiltration and functionality. Mechanistically, TP63 and STAT1 mutually suppress each other to regulate the IFNγ signaling by co-occupying and co-regulating their own promoters and enhancers. Together, our findings elucidate a tumor-extrinsic function of TP63 in promoting immune evasion of SCC cells. Over-expression of TP63 may serve as a biomarker predicting the outcome of SCC patients treated with ICB therapy, and targeting TP63/STAT/IFNγ axis may enhance the efficacy of ICB therapy for this deadly cancer.

Establishing mouse and human oral esophageal organoids to investigate the tumor immune response.

Organoid culture systems are very powerful models that recapitulate in vivo organ development and disease pathogenesis, offering great promise in basic research, drug screening and precision medicine. However, the application of organoids derived from patients with cancer to immunotherapeutic research is a relatively untapped area. Esophageal cancer is one of the most lethal malignancies worldwide, including two major pathological subtypes: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma. ESCC shares many biological and genomic features with oral squamous cell cancers. Herein, we provide a versatile protocol for the establishment and maintenance of oral and esophageal organoid cultures derived from both murine and human samples. We describe culture conditions for organoids derived from normal tongue, esophagus and gastroesophageal junction, esophageal cancer and Barrett's esophagus. In addition, we establish an ex vivo model by co-culturing patient tumor-derived organoids and autologous CD8+ T lymphocytes to assess CD8+ T cell-mediated tumor killing. Our protocol can also be modified for organoid establishment from other squamous epithelia and carcinomas. The co-culture model can serve as a template for studies of other tumor-immune cell interactions and the efficacy of immune checkpoint blockade therapy.

Zona pellucida family genes in Chinese pond turtle: identification, expression profiles, and role in the spermatozoa acrosome reaction†.

The zona pellucida (ZP) is an extracellular matrix that surrounds all vertebrate eggs, and it is involved in fertilization and species-specific recognition. Numerous in-depth studies of the ZP proteins of mammals, birds, amphibians, and fishes have been conducted, but systematic investigation of the ZP family genes and their role during fertilization in reptiles has not been reported to date. In this study, we identified six turtle ZP (Tu-ZP) gene subfamilies (Tu-ZP1, Tu-ZP2, Tu-ZP3, Tu-ZP4, Tu-ZPD, and Tu-ZPAX) based on whole genome sequence data from Mauremys reevesii. We found that Tu-ZP4 had large segmental duplication and was distributed on three chromosomes, and we also detected gene duplication in the other Tu-ZP genes. To evaluate the role of Tu-ZP proteins in sperm-egg binding, we assessed the expression pattern of these Tu-ZP proteins and their ability to induce the spermatozoa acrosome reaction in M. reevesii. In conclusion, this is the first report of the existence of gene duplication of Tu-ZP genes and that Tu-ZP2, Tu-ZP3, and Tu-ZPD can induce acrosome exocytosis of spermatogenesis in the reptile.

[Source Analysis and Ecological Risk Assessment of Heavy Metals in the Arable Soil at the Geological High Background, Based on the Township Scale].

Due to the extensive development of carbonate rocks in southwest China, heavy metals are naturally occurring elements that have high natural background levels in the environment. Therefore, it is important to conduct ecological risk assessments and identify potential sources of heavy metals in the geological high background area. Based on the township scale, a total of 307 surface soil samples were collected in Qinglong Town, Fengjie County, Chongqing. The concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn and pH were analyzed and determined. The spatial distribution and source of soil heavy metals were discussed using the geostatistical analysis and an absolute principal component score-multiple linear regression (APCS-MLR) model in the studied area. The results showed that the average values of seven heavy metals (As, Cd, Cr, Cu, Hg, Ni, and Zn) in the arable soil exceeded the background values of Chongqing, and the cumulative effect of Cd and As was obvious. The concentrations of Cd significantly exceededthe screening values in The Risk Control Standard for Soil Environmental Quality and Soil Pollution in Agricultural Land (GB 15618-2018), with the over-standard rates of 52.12%. The spatial characteristics of soil heavy metal contents exhibited a pattern of high in the south and low in the north. PCA and APCS-MLR modeling revealed that the contributions of natural sources to Cr, Cu, Ni, and Zn were 86.62%, 64.34%, 76.44%, and 85.46%, respectively. As, Pb, and Hg mainly derived from industrial activities, which accounted for 74.63%, 61.90%, and 73.49%, respectively, and Cd was affected by both natural sources and industrial activities (accounting for 47.74% and 39.56% of the total Cd content, respectively). The evaluation of the soil by the Nemerow comprehensive index (P) showed that Cd pollution was relatively serious, accounting for 27.04% of soil pollution. The potential ecological hazard index showed that Cd and Hg were the main ecological hazard elements, and the distribution range of RI was 51.77 to 2228, indicating mainly mild and moderate risks, and the moderate and above risk areas in the study area were mainly located around the southern industrial source area. Altogether, our results revealed that in the study area, the heavy metal pollution was mainly caused by industrial activities, and the heavy metal pollution caused by geological background was mainly light to moderate. In conclusion, the medium and above risk areas in the study area were mainly caused by mineral and industrial activities, whereas the heavy metal pollution caused by geological background was mainly light to moderate pollution.

CTNNBL1 restricts HIV-1 replication by suppressing viral DNA integration into the cell genome.

Retroviral integration is mediated by a unique enzymatic process shared by all retroviruses and retrotransposons. During integration, double-stranded linear viral DNA is inserted into the host genome in a process catalyzed by viral-encoded integrase (IN). However, host cell defenses against HIV-1 integration are not clear. This study identifies ß-catenin-like protein 1 (CTNNBL1) as a potent inhibitor of HIV-1 integration via association with viral-encoded integrase (IN) and its cofactor, lens epithelium-derived growth factor/p75. CTNNBL1 overexpression blocks HIV-1 integration and inhibits viral replication, whereas CTNNBL1 depletion significantly upregulates HIV-1 integration into the genome of various target cells. Further, CTNNBL1 expression is downregulated in CD4+ T cells by activation, and CTNNBL1 depletion also facilitates HIV-1 integration in resting CD4+ T cells. Thus, host cells may employ CTNNBL1 to inhibit HIV-1 integration into the genome. This finding suggests a strategy for the treatment of HIV infections.

Comparison of Five Triage Tools for Identifying Mortality Risk and Injury Severity of Multiple Trauma Patients Admitted to the Emergency Department in the Daytime and Nighttime: A Retrospective Study.

Effective triage tools are indispensable for doctors to make a prompt decision for the treatment of multiple trauma patients in emergency departments (EDs). The Modified Early Warning Score (MEWS), National Early Warning Score (NEWS), standardized early warning score (SEWS), Modified Rapid Emergency Medicine Score (mREMS), and Revised Trauma Score (RTS) are five common triage tools proposed for trauma management. However, few studies have compared these tools in a multiple trauma cohort and investigated the influence of nighttime admission on the performance of these tools. This retrospective study was aimed at evaluating and comparing the performance of MEWS, NEWS, SEWS, mREMS, and RTS for identifying the mortality risk and trauma severity of patients with multiple trauma admitted to the ED during the daytime and nighttime. Retrospective data were collected from the medical records of patients with multiple trauma admitted in the daytime or nighttime to calculate scores for each triage tool. Logistic regression analysis was conducted on each triage tool for identifying in-hospital mortality and severe trauma (injury severity score > 15) in the daytime and nighttime. The performance of the tools was evaluated and compared by calculating area under the receiver operating characteristic curve (AUROC) of the retrospective logistic model of each tool. We collected data for 1,818 admissions, including 1,070 daytime and 748 nighttime admissions. A comparison of performance for identifying in-hospital mortality between daytime and nighttime yielded the following results (AUROC): MEWS (0.95 vs. 0.93, p = 0.384), NEWS (0.95 vs. 0.94, p = 0.708), SEWS (0.95 vs. 0.94, p = 0.683), mREMS (0.94 vs. 0.92, p = 0.286), and RTS (0.93 vs. 0.93, p = 0.87). Similarly, a comparison of performance for identifying trauma severity between daytime and nighttime yielded the following results (AUROC): MEWS (0.78 vs. 0.78, p = 0.95), NEWS (0.8 vs. 0.8, p = 0.885), SEWS (0.78 vs. 0.78, p = 0.818), mREMS (0.75 vs. 0.69, p = 0.019), and RTS (0.75 vs. 0.74, p = 0.619). All five scores are excellent triage tools (AUROC ≥ 0.9) for identifying in-hospital mortality for both daytime and nighttime admissions. However, they have only moderate effectiveness (AUROC < 0.9) at identifying severe trauma. The NEWS is the best triage tool for identifying severe trauma for both daytime and nighttime admissions. The MEWS, NEWS, SEWS, and RTS exhibited no significant differences in performance for identifying in-hospital mortality or severe trauma during the daytime or nighttime. However, the mREMS was better at identifying severe trauma during the daytime.

Chromosome-level genome assembly of the Chinese three-keeled pond turtle (Mauremys reevesii) provides insights into freshwater adaptation.

Mauremys reevesii is an endangered freshwater turtle that symbolizes longevity in Chinese culture. Despite its importance, genetic studies of this species remain limited, with no genomic sequence reported to date. Here, we report a high-quality, chromosome-level genomic sequence of M. reevesii obtained using a combination of Nanopore and Hi-C sequencing technologies. The 2.37 Gb M. reevesii genome was assembled from a total of ~226.80 Gb of Nanopore sequencing data. The M. reevesii genome contig N50 is 34.73 Mb, the highest value in published turtle genomes. In total, 18,238 genes were functionally annotated. The contigs were clustered and ordered onto 27 pseudochromosomes covering ~96.55% of the genome assembled with Hi-C data. To explore genome evolution, synteny analysis was performed between M. reevesii (freshwater turtle) and Gopherus evgoodei (terrestrial turtle) genomes. In general, each chromosome of M. reevesii corresponded to one chromosome of Gopherus evgoodei, but some interchromosomal rearrangements occurred between the two species based on the assembled genomes. These interchromosomal rearrangements were further confirmed by mapping of the long-read nanopore data to the assembly. The reconstructed demographic history showed varied effective population size among freshwater, marine and terrestrial turtles. We also discovered expansion of genes related to the innate immune system in M. reevesii that may provide defence against freshwater pathogens. The high-quality genomic sequence provides a valuable genetic resource for further studies of genetics and genome evolution in turtles.

The interaction between Tu-Izumo1 and Tu-JUNO is involved in turtles hybridization.

The specificity of sperm-egg recognition is crucial to species independence, and two proteins (Izumo1 and JUNO) are essential for gamete adhesion/fusion in mammals. However, hybridization, which is very common in turtles, also requires specific recognition of sperm-egg binding proteins. In this study, we discovered that natural selection plays an important role in the codon usage bias of Tu-Izumo1 and Tu-JUNO. Positively selected sites and co-evolutionary analyses between Tu-Izumo1 and Tu-JUNO have been previously reported, and we confirm these results in a larger analysis containing 25 turtle species. We also showed that Tu-JUNO is expressed on the oocyte surface and that Tu-Izumo1 and Tu-JUNO interact with each other directly in different species hybridization combinations. Co-immunization assays revealed that this interaction is evolutionarily conserved in turtles. The results of avidity-based extracellular interaction screening between Tu-Izumo1 and Tu-JUNO for sperm-oocyte binding pairs (both within and across species) likely suggest that the interaction force between Izumo1 and JUNO has a certain correlation in whether the turtles can hybridize. Our results lay a theoretical foundation for the subsequent development of techniques to detect whether different turtle species can interbreed, which would provide the molecular basis for breeding management and species protection of turtles.

Morphological mechanism allowing a parasitic leech, Ozobranchus jantseanus (Rhynchobdellida: Ozobranchidae), to survive in ultra-low temperatures.

Ozobranchus jantseanus is the largest metazoan known to survive in liquid nitrogen without pretreatment to date; however, the mechanism underlying this tolerance remains unclear. In this study, the first analyses of histological and morphological changes in normal, frozen, and dehydrated states were performed. Adults survived after direct placement in liquid nitrogen for 96 h, with a survival rate of approximately 86.7%. The leech could withstand rapid desiccation and its survival rate after rehydration was 100% when its water loss was below about 84.8%. After freezing, desiccation, and ethanol dehydration, the leech immediately formed a hemispherical shape. Particularly during drying, an obvious transparent glass-like substance was observed on surface. Scanning electron microscopy revealed many pores on the surface of the posterior sucker, creating a sponge-like structure, which may help to rapidly expel water, and a hemispherical shape may protect the internal organs by contraction and folding reconstruction in the anterior-posterior direction. A substantial amount of mucopolysaccharides on the surface and acid cells and collagen fibers in the body, all of which contained substantial polysaccharides, may play a key protective role during freezing. Our results indicate that the resistance of leeches to ultra-low temperatures can be explained by cryoprotective dehydration/vitrification strategies. This article has an associated First Person interview with the first author of the paper.

[Characteristics of Cadmium Enrichment and Pollution Evaluation of a Soil-Crop System in a Typical Karst Area].

In order to study the characteristics and factors influencing Cd accumulation in surface soils and crops in karst areas, and to provide a theoretical basis for safe land use, 360 surface soil samples, 7 deep soil samples, and 85 rice samples were collected from central Qianjiang District, Chongqing. The samples and 73 corn samples (corresponding to root-zone soil samples), were analysed to determine the content of Cd, TFe2 O3, Mn, organic matter (Corg), Se, and pH. Based on geostatistical analyses, the spatial distribution and Cd enrichment of the surface soils were determined and a safety evaluation for the soil and crops was carried out. The results showed that the spatial distribution of Cd in the surface soil was uneven, with the surface layer showing significant enrichment. This pattern was controlled by the soil parent material and human activities. The enrichment of surface layer was mainly affected by iron manganese oxides and organic matter (Corg). Soil Cd was mainly found at 'non-polluted' and 'lightly polluted' levels, although some areas present strong ecological risks. The main contaminated area occurs in association with Permian strata, demonstrating a geological control on soil Cd pollution. Slight-to-severe Cd pollution was identified in bulk crops; the recommended daily consumption limit for rice is 0.87 kg·d-1 and corn is 1.53 kg·d-1. The bioavailability of Cd is affected by soil pH and Se content. Under acidic conditions, Cd bioavailability is high, and crops in areas with high soil Se are safer. It is recommended that crops with low Cd accumulation are planted in the Permian outcrop area of Shuitian Township, or alternatively, soil pH should be adjusted to control the risk of Cd pollution and ensure safe land use. In addition, planting crops in areas with high soil Se content is preferable.

[Distribution Characteristics of Selenium in a Soil-Crop System and the Threshold of Selenium-Rich Soils].

In order to determine the distribution characteristics of Se in soil-crop systems, we carried out a study on the Se-rich soil threshold by collecting 8789 surface soils and 155 deep soils in the Qianjiang District of Chongqing City, China, and 141 corn seeds and 159 rice seeds (simultaneously collecting 141 and 159 corresponding root soil samples, respectively). We then analyzed the Se content, organic matter, S, Mn, TFe2O3, Al2O3, and K2O in soils and crops, and soil pH. We also analyzed the surface layer using geostatistical methods and the distribution characteristics of Se in deep soils using multiple regression analysis to study the factors influencing the bioavailability of Se. Based on the contents of each component of root soil and the Se contents of crops, the Se rich threshold was examined. The results showed that the high-Se soils in the study area account for 32.72% of the total area; the distribution of Se contents in the surface and deep soils is mainly controlled by the parent material, the source of soil Se is stable, and the surface enrichment is obvious. The Se-rich rates of corn and rice were 75.35% and 46.81%, respectively, and soil organic matter and S content will limit the bioavailability of Se. If the planted crop is corn, it is recommended to use 0.3 mg·kg-1 as the Se-rich soil threshold; if the planted crop is rice, when the soil pH is ≤ 7.5, it is recommended to use 0.3 mg·kg-1 as the Se-rich soil threshold, while at a soil pH>7.5, it is recommended to use 0.4 mg·kg-1 as the threshold. Similarly, if other large crops are planted in the study area, this method can also be used to carry out a study on the proposed Se-rich soil threshold.

Characterization of deoxyribonucleic methylation and transcript abundance of sex-related genes during tempera ture-dependent sex determination in Mauremys reevesii†.

A number of genes relevant for sex determination have been found in species with temperature-dependent sex determination. Epigenetics play a key role in sex determination, but characterization of deoxyribonucleic acid methylation of sex-related genes on temperature-dependent sex determination remains unclear. Mauremys reevesii is a typical species with temperature-dependent sex determination. In this study, we analyzed the Cytosine Guanine (CpG) methylation status of the proximal promoters, the messenger ribonucleic acid expression patterns and the correlation between methylation and expression levels of Aromatase, Forkhead box protein L2, Doublesex and mab3-related transcription factor 1, sex-determining region on Y chromosome-box 9, and anti-Müllerian hormone, which are key genes in sex determination in other species. We also analyzed the expression level of genes that encode enzymes involved in methylation and demethylation. The expression levels of Aromatase and Forkhead box protein L2 at the female producing temperature were higher than those at the male producing temperature; the expression levels of Doublesex and mab3-related transcription factor 1, sex-determining region on Y chromosome-box 9, and anti-Müllerian hormone were higher at MPT. The expression of some genes involved in methylation and demethylation is significantly different between male producing temperature and female producing temperature. The expression of messenger ribonucleic acid of genes involved in deoxyribonucleic acid methylation and demethylation affected by temperature, together with other factors, may change the methylation level of the regulatory regions of sex-related genes, which may further lead to temperature-specific expression of sex-related genes, and eventually affect the differentiation of the gonads.

Next-Generation Genome Sequencing of Sedum plumbizincicola Sheds Light on the Structural Evolution of Plastid rRNA Operon and Phylogenetic Implications within Saxifragales.

The genus Sedum, with about 470 recognized species, is classified in the family Crassulaceae of the order Saxifragales. Phylogenetic relationships within the Saxifragales are still unresolved and controversial. In this study, the plastome of S. plumbizincicola was firstly presented, with a focus on the structural analysis of rrn operon and phylogenetic implications within the order Saxifragaceae. The assembled complete plastome of S. plumbizincicola is 149,397 bp in size, with a typical circular, double-stranded, and quadripartite structure of angiosperms. It contains 133 genes, including 85 protein-coding genes (PCGs), 36 tRNA genes, 8 rRNA genes, and four pseudogenes (one ycf1, one rps19, and two ycf15). The predicted secondary structure of S. plumbizincicola 16S rRNA includes three main domains organized in 74 helices. Further, our results confirm that 4.5S rRNA of higher plants is associated with fragmentation of 23S rRNA progenitor. Notably, we also found the sequence of putative rrn5 promoter has some evolutionary implications within the order Saxifragales. Moreover, our phylogenetic analyses suggested that S. plumbizincicola had a closer relationship with S. sarmentosum than S. oryzifolium, and supported the taxonomic revision of Phedimus. Our findings of the present study will be useful for further investigation of the evolution of plastid rRNA operon and phylogenetic relationships within Saxifragales.

Membrane metalloprotease TRABD2A restricts HIV-1 progeny production in resting CD4+ T cells by degrading viral Gag polyprotein.

Resting CD4+ T cells are highly resistant to the production of human immunodeficiency virus type 1 (HIV-1). However, the mechanism by which resting CD4+ T cells restrict such production in the late viral replication phase of infection has remained unclear. In this study, we found that the cell membrane metalloprotease TRAB domain-containing protein 2A (TRABD2A) inhibited this production in resting CD4+ T cells by degrading the virion structural precursor polyprotein Gag at the plasma membrane. Depletion or inhibition of metalloprotease activity by TRABD2A profoundly enhanced HIV-1 production in resting CD4+ T cells. TRABD2A expression was much higher in resting CD4+ T cells than in activated CD4+ T cells and was considerably reduced by T cell activation. Moreover, reexpressing TRABD2A reinforced the resistance of activated CD4+ T cells to the production of HIV-1 progeny. Collectively, these results elucidate the molecular mechanism employed by resting CD4+ T cells to potently restrict the assembly and production of HIV-1 progeny.

Transcriptome sequencing and comparative analysis of adult ovary and testis identify potential gonadal maintenance-related genes in Mauremys reevesii with temperature-dependent sex determination.

Mauremys reevesii is a classical organism with temperature-dependent sex determination (TSD). Gonad development in early life has recently received considerable attention but gonadal maintenance after sex differentiation in turtles with TSD remains a mystery. In this study, we sequenced the transcriptomes for the adult testis and ovary using RNA-seq, and 36,221 transcripts were identified. In total, 1,594 differentially expressed genes (DEGs) were identified where 756 DEGs were upregulated in the testis and 838 DEGs were upregulated in the ovary. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis suggested that the TGF-beta signaling pathway and Hedgehog signaling pathway have important roles in testis maintenance and spermatogenesis, whereas the Hippo signaling pathway and Wnt signaling pathway are likely to participate in ovary maintenance. We determined the existence of antagonistic networks containing significant specific-expressed genes and pathways related to gonadal maintenance and gametogenesis in the adult gonads of M. reevesii. The candidate gene Fibronectin type 3 and ankyrin repeat domains 1 (FANK1) might be involved with the regulation of testis spermatogenesis.

Detecting coevolution of positively selected in turtles sperm-egg fusion proteins.

Physically interacting sperm-egg proteins have been identified using gene-modified animals in some mammal species. Three proteins are essential for sperm-egg binding: Izumo1 on the sperm surface, and JUNO and CD9 on the egg surface. Most proteins linked to reproductive function evolve rapidly among species by positive selection, and have correlated evolutionary rates to compensate for changes on both the sperm and egg. Up to now, interactions between sperm and egg proteins have not been identified in non-mammalian vertebrates, such as turtles that have interspecific hybrids that can produce surviving F1 generations. To explore the potential physical interactions of sperm-egg proteins in turtle species, the coding region of Izumo1, JUNO, and CD9 homologous genes (named Tu-Izumo1, Tu-JUNO, and Tu-CD9) in six turtle species (Mauremys reevesii, M. mutica, M. sinensis, Cistoclemmys flavomarginata, Platysternon megacephalum and Chrysemys picta bellii) were identified, amplified, and sequenced, and tissue-specific expression was analyzed in M. reevesii. We constructed phylogenetic trees and analyzed the signatures of coevolution between sperm-egg protein pairs using MirrorTree Server and linear regression methods. The results showed that Tu-Izumo1, Tu-JUNO, and Tu-CD9 proteins have correlated evolutionary rates, and that the area where Tu-Izumo1 interacts with Tu-JUNO has only one positive selection site in some turtle species. These results suggest there is a potential interaction between Tu-Izumo1 and Tu-JUNO among turtles that can interbreed, and that a significantly lower positive selection in the interaction region may be one of the reasons why turtle hybrids are so common. Further studies are required to uncover Tu-Izumo1, Tu-JUNO and Tu-CD9 protein biological functions during gamete fusion.

Single-molecule long-read transcriptome profiling of Platysternon megacephalum mitochondrial genome with gene rearrangement and control region duplication.

Platysternon megacephalum is the sole living representative of the poorly studied turtle lineage Platysternidae. Their mitochondrial genome has been subject to gene rearrangement and control region duplication, resulting in a unique mitochondrial gene order in vertebrates. In this study, we sequenced the first full-length turtle (P. megacephalum) liver transcriptome using single-molecule real-time sequencing to study the transcriptional mechanisms of its mitochondrial genome. ND5 and ND6 anti-sense (ND6AS) forms a single transcript with the same expression in the human mitochondrial genome, but here we demonstrated differential expression of the rearranged ND5 and ND6AS genes in P. megacephalum. And some polycistronic transcripts were also reported in this study. Notably, we detected some novel long non-coding RNAs with alternative polyadenylation from the duplicated control region, and a novel ND6AS transcript composed of a long non-coding sequence, ND6AS, and tRNA-GluAS. These results provide the first description of a mtDNA transcriptome with gene rearrangement and control region duplication. These findings further our understanding of the fundamental concepts of mitochondrial gene transcription and RNA processing, and provide a new insight into the mechanism of transcription regulation of the mitochondrial genome.

The Per-1 Short Isoform Inhibits de novo HIV-1 Transcription in Resting CD4+ T-cells.

BACKGROUND: Understanding of the restriction of HIV-1 transcription in resting CD4+ Tcells is critical to find a cure for AIDS. Although many negative factors causing HIV-1 transcription blockage in resting CD4+ T-cells have been found, there are still unknown mechanisms to explore. OBJECTIVE: To explore the mechanism for the suppression of de novo HIV-1 transcription in resting CD4+ T-cells. METHODS: In this study, a short isoform of Per-1 expression plasmid was transfected into 293T cells with or without Tat's presence to identify Per-1 as a negative regulator for HIV-1 transcription. Silencing of Per-1 was conducted in resting CD4+ T-cells or monocyte-derived macrophages (MDMs) to evaluate the antiviral activity of Per-1. Additionally, we analyzed the correlation between Per-1 expression and viral loads in vivo, and silenced Per-1 by siRNA technology to investigate the potential anti-HIV-1 roles of Per-1 in vivo in untreated HIV-1-infected individuals. RESULTS: We found that short isoform Per-1 can restrict HIV-1 replication and Tat ameliorates this inhibitory effect. Silencing of Per-1 could upregulate HIV-1 transcription both in resting CD4+ Tcells and MDMs. Moreover, Per-1 expression is inversely correlated with viral loads in Rapid progressors (RPs) in vivo. CONCLUSION: These data together suggest that Per-1 is a novel negative regulator of HIV-1 transcription. This restrictive activity of Per-1 to HIV-1 replication may contribute to HIV-1 latency in resting CD4+ T-cells.