Functional Switch Research Articles

Abstract PO-055: Global immune characterization of hepatocellular carcinoma identifies macrophage and T cell subsets related to disease progression

Abstract Diverse immune cells in the tumor microenvironment form a complex ecosystem, but our knowledge of their heterogeneity and dynamics within hepatocellular carcinoma (HCC) remains poorly understood. We assess the plasticity and phenotypes of immune cells within HCC microenvironment at single-cell level. We performed single-cell RNA sequencing on 41,698 immune cells from 7 pairs of HCC tumors and nontumor liver tissues. We combined bioinformatic analyses, flow cytometry and multiplex immunohistochemistry to assess the heterogeneity of different immune subsets in functional characteristics, transcriptional regulation, phenotypic switching, and interactions. We identified 29 immune subsets of myeloid cells, NK cells, and lymphocytes with unique transcriptomic profiles in HCC. A highly complex immunological network was shaped by immune subsets with diverse characteristics that can transit among different states and mutually engage through intercellular communication. Notably, we identified a subset of M2 macrophage with high expression of CCL18 and transcription factor CREM that was enriched in advanced HCC patients, and potentially participated in tumor progression. We also detected a new subset of activated CD8+ T cells highly expressing XCL1 that correlated with better patient survival, and possibly enhanced anti-tumor response. Meanwhile, distinct transcriptomic signatures, cytotoxic phenotypes and evolution trajectory of effector CD8+ T cells from early-stage to advanced HCC were also identified. Our study presents a comprehensive insight for dissecting the immune microenvironment in HCC and highlights novel macrophage and T cell subsets that could be further exploited in future immunotherapy. Citation Format: Guohe Song, Yang Shi, Youpei Lin, Juan Zhang, Shuaixi Yang, Qiang Gao. Global immune characterization of hepatocellular carcinoma identifies macrophage and T cell subsets related to disease progression [abstract]. In: Proceedings of the AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact; 2020 Sep 17-18. Philadelphia (PA): AACR; Cancer Res 2020;80(21 Suppl):Abstract nr PO-055.

Destabilisation of the structure of transthyretin is driven by Ca2+

Tetrameric transthyretin (TTR) transports thyroid hormones and retinol in plasma and cerebrospinal fluid and performs protective functions under stress conditions. Ageing and mutations result in TTR destabilisation and the formation of the amyloid deposits that dysregulate Ca2+ homeostasis. Our aim was to determine whether Ca2+ affects the structural stability of TTR. We show, using multiple techniques, that Ca2+ does not induce prevalent TTR dissociation and/or oligomerisation. However, in the presence of Ca2+, TTR exhibits altered conformational flexibility and different interactions with the solvent molecules. These structural changes lead to the formation of the sub-populations of non-native TTR conformers and to the destabilisation of the structure of TTR. Moreover, the sub-population of TTR molecules undergoes fragmentation that is augmented by Ca2+. We postulate that Ca2+ constitutes the structural and functional switch between the native and non-native forms of TTR, and therefore tip the balance towards age-dependent pathological calcification.

Chain-Selective Isotopic Labeling of the Heterodimeric Type III Secretion Chaperone, Scc4:Scc1, Reveals the Total Structural Rearrangement of the Chlamydia trachomatis Bi-Functional Protein, Scc4.

Scc4 is an unusual bi-functional protein from Chlamydia trachomatis (CT) that functions as a type III secretion system (T3SS) chaperone and an RNA polymerase (RNAP)-binding protein. Both functions require interactions with protein partners during specific stages of the CT developmental cycle. As a T3SS chaperone, Scc4 binds Scc1 during the late stage of development to form a heterodimer complex, which chaperones the essential virulence effector, CopN. During the early-middle stage of development, Scc4 regulates T3SS gene expression by binding the σ66-containing RNAP holoenzyme. In order to study the structure and association mechanism of the Scc4:Scc1 T3SS chaperone complex using nuclear magnetic resonance (NMR) spectroscopy, we developed an approach to selectively label each chain of the Scc4:Scc1 complex with the 15N-isotope. The approach allowed one protein to be visible in the NMR spectrum at a time, which greatly reduced resonance overlap and permitted comparison of the backbone structures of free and bound Scc4. 1H,15N-heteronuclear single quantum coherence spectra of the 15N-Scc4:Scc1 and Scc4:15N-Scc1 complexes showed a total structural rearrangement of Scc4 upon binding Scc1 and a dynamic region isolated to Scc1, respectively. Development of the chain-selective labeling approach revealed that the association of Scc4 and Scc1 requires partial denaturation of Scc1 to form the high affinity complex, while low affinity interactions occurred between the isolated proteins under non-denaturing conditions. These results provide new models for Scc4′s functional switching mechanism and Scc4:Scc1 association in CT.

Ultra-short-term combined prediction approach based on kernel function switch mechanism

A novel combined prediction approach including nonlinear data analysis, data decomposition and reconstruction, and parameter optimization is proposed to improve the accuracy of ultra-short-term wind power prediction. The distance correlation coefficient (DCC) is applied to analyze the non-linear correlation between wind power and numerical weather prediction (NWP) data. The original wind power is decomposed by the complete ensemble empirical mode decomposition with adaptive noise (CEEMDAN) technique to eliminate the noise of the data, and then the decomposed wind power is reconstructed into new subsequences by weight permutation entropy (WPE) according to time series complexity. The least squares support vector machine (LSSVM) prediction model based on the kernel function switch mechanism is developed by the use of the new subsequence and key factor as input variables. Furthermore, an improved cuckoo search (ICS) strategy is established to optimize the penalty factor and kernel function parameters of the prediction model. The validation of the proposed model is tested with data from four actual wind farms at a large wind farm base in Northeast China. The results show that the proposed approach is effective with higher accuracy compared with other prediction approaches.

Phosphorylation at S548 as a Functional Switch of Sterile Alpha and TIR Motif-Containing 1 in Cerebral Ischemia/Reperfusion Injury in Rats.

Sterile alpha and Toll/interleukin-1 receptor motif-containing 1 (SARM1) is a pro-degenerative molecule in Wallerian degeneration, which is mainly expressed in brain/neurons and colocalized with mitochondria and microtubules. The aim of this study was to determine the role of SARM1 in cerebral ischemia/reperfusion (I/R) injury and the underlying mechanisms. In vivo, a middle cerebral artery occlusion/reperfusion (MCAO/R) model in adult male Sprague Dawley rats (250-300 g) was established, and in vitro, cultured primary neurons were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to imitate I/R injury. Overexpression lentiviruses encoding wild-type SARM1 and SARM1 with serine 548 alanine mutation (S548A) were constructed and administered to rats by intra-penumbral injection. First, the potential role of SARM1 in cerebral I/R injury was confirmed by the increased protein levels of SARM1 within penumbra tissue, especially in neurons. Second, there was an increase in the phosphorylation ratio of p-SARM1(S548)/SARM1 at 2 h after MCAO/R. Third, on the basis of site-specific mutagenesis, we identified S548 as a key site for SARM1 phosphorylation in I/R conditions. Fourth, SARM1 (S548A) overexpression reduced infarct size, neuronal death, and neurobehavioral dysfunction, while wild-type SARM1 overexpression had the opposite effects. Finally, we found that SARM1 phosphorylation at the S548 site switched SARM1 function from promoting mitochondrial transport to inhibiting mitochondrial transport along axons after I/R injury. Restriction of SARM1 phosphorylation at S548 may be a promising intervention target for I/R injury; thus, exogenous administration of SARM1 (S548A) may be a novel strategy for improving neurological outcomes.

Genome-wide transcriptome and proteome profiles indicate an active role of alternative splicing during de-etiolation of maize seedlings

AS events affect genes encoding protein domain composition and make the single gene produce more proteins with a certain number of genes to satisfy the establishment of photosynthesis during de-etiolation. The drastic switch from skotomorphogenic to photomorphogenic development is an excellent system to elucidate rapid developmental responses to environmental stimuli in plants. To decipher the effects of different light wavelengths on de-etiolation, we illuminated etiolated maize seedlings with blue, red, blue-red mixed and white light, respectively. We found that blue light alone has the strongest effect on photomorphogenesis and that this effect can be attributed to the higher number and expression levels of photosynthesis and chlorosynthesis proteins. Deep sequencing-based transcriptome analysis revealed gene expression changes under different light treatments and a genome-wide alteration in alternative splicing (AS) profiles. We discovered 41,188 novel transcript isoforms for annotated genes, which increases the percentage of multi-exon genes with AS to 63% in maize. We provide peptide support for all defined types of AS, especially retained introns. Further in silico prediction revealed that 58.2% of retained introns have changes in domains compared with their most similar annotated protein isoform. This suggests that AS acts as a protein function switch allowing rapid light response through the addition or removal of functional domains. The richness of novel transcripts and protein isoforms also demonstrates the potential and importance of integrating proteomics into genome annotation in maize.

A metabolite binding protein moonlights as a bile-responsive chaperone.

Bile salts are secreted into the gastrointestinal tract to aid in the absorption of lipids. In addition, bile salts show potent antimicrobial activity in part by mediating bacterial protein unfolding and aggregation. Here, using a protein folding sensor, we made the surprising discovery that the Escherichia coli periplasmic glycerol-3-phosphate (G3P)-binding protein UgpB can serve, in the absence of its substrate, as a potent molecular chaperone that exhibits anti-aggregation activity against bile salt-induced protein aggregation. The substrate G3P, which is known to accumulate in the later compartments of the digestive system, triggers a functional switch between UgpB's activity as a molecular chaperone and its activity as a G3P transporter. A UgpB mutant unable to bind G3P is constitutively active as a chaperone, and its crystal structure shows that it contains a deep surface groove absent in the G3P-bound wild-type UgpB. Our work illustrates how evolution may be able to convert threats into signals that first activate and then inactivate a chaperone at the protein level in a manner that bypasses the need for ATP.

Structure optimization of positive allosteric modulators of GABAB receptors led to the unexpected discovery of antagonists/potential negative allosteric modulators

Positive allosteric modulators (PAMs) of GABAB receptor represent an interesting alternative to receptor agonists such as baclofen, as they act on the receptor in a more physiological way and thus are devoid of the side effects typically exerted by the agonists. Based on our interest in the identification of new GABAB receptor PAMs, we followed a merging approach to design new chemotypes starting from selected active compounds, such as GS39783, rac-BHFF, and BHF177, and we ended up with the synthesis of four different classes of compounds. The new compounds were tested alone or in the presence of 10 µM GABA using [35S]GTPγS binding assay to assess their functionality at the receptor. Unexpectedly, a number of them significantly inhibited GABA-stimulated GTPγS binding thus revealing a functional switch with respect to the prototype molecules. Further studies on selected compounds will clarify if they act as negative modulators of the receptor or, instead, as antagonists at the orthosteric binding site.

Advances of RRAM Devices: Resistive Switching Mechanisms, Materials and Bionic Synaptic Application.

Resistive random access memory (RRAM) devices are receiving increasing extensive attention due to their enhanced properties such as fast operation speed, simple device structure, low power consumption, good scalability potential and so on, and are currently considered to be one of the next-generation alternatives to traditional memory. In this review, an overview of RRAM devices is demonstrated in terms of thin film materials investigation on electrode and function layer, switching mechanisms and artificial intelligence applications. Compared with the well-developed application of inorganic thin film materials (oxides, solid electrolyte and two-dimensional (2D) materials) in RRAM devices, organic thin film materials (biological and polymer materials) application is considered to be the candidate with significant potential. The performance of RRAM devices is closely related to the investigation of switching mechanisms in this review, including thermal-chemical mechanism (TCM), valance change mechanism (VCM) and electrochemical metallization (ECM). Finally, the bionic synaptic application of RRAM devices is under intensive consideration, its main characteristics such as potentiation/depression response, short-/long-term plasticity (STP/LTP), transition from short-term memory to long-term memory (STM to LTM) and spike-time-dependent plasticity (STDP) reveal the great potential of RRAM devices in the field of neuromorphic application.

Assessment of Hybrid RANS/LES Models in Heat and Fluid Flows around Staggered Pin-Fin Arrays

In the present work, the three-dimensional heat and fluid flows around staggered pin-fin arrays are predicted using two hybrid RANS/LES models (an improved delayed detached eddy simulation (IDDES) model and a stress-blended eddy simulation (SBES) model), and one transitional unsteady Reynolds averaged Navier-Stokes (URANS) model, called k-ω SSTLM. The periodic segment geometry with a total of nine pins is considered with a channel height of 2D and a distance of 2.5D between each pin. The corresponding Reynolds number based on the pin diameter and the maximum velocity between pins is 10,000. The two hybrid RANS/LES results show the superior prediction of the mean velocity profiles around the pins, pressure distributions on the pin wall, and Nusselt number distributions. However, the transitional model, k-ω SSTLM, shows large discrepancies except in front of the pins where the flow is not fully developed. The vortical structures are well resolved by the two hybrid RANS/LES models. The SBES model is particularly adept at capturing the 3-D vortex structures after the pins. The effects of the blending function switching between RANS and LES mode of the two hybrid RANS/LES models are also investigated.

Polymer-chlorella cells conjugating with aggregation-induced functionality switch towards hydrogen evolution

The manipulation of physicochemical and biological properties of the encapsulated cells by cell surface engineering technique is now attracting more and more researchers’ interest. In this study, we showed a mild and highly efficient way to build a type of polymer-living cell conjugates based on photoinduced electron transfer-reversible addition-fragmentation chain-transfer polymerization (PET-RAFT). The polymerization could proceed at room temperature under the blue light initiating and showed very less influence on the activity of the living cells. Significantly, the constructed positively charged polymer-living cell conjugates could aggregate with the negatively charged native Chlorella pyrenoidosa (CP) cell to form a microscale aggregate which then allowed a functionality switching of the encapsulated CP cells from the normal photosynthetic O2 towards H2 production with an average rate of 0.26 μmol H2h−1 (mg chlorophyll)−1 for 42 h. It is then anticipated that the method we show could contribute an effective way for generating different functionalized living cell-polymer hybrid structures on demand, and the demonstrated aggregation-induced functionality switching of the CP cells is also anticipated to inspire more applications of the cell-based materials towards biology green energy.

RNA editing mediates the functional switch of COPA in a novel mechanism of hepatocarcinogenesis

RNA editing introduces nucleotide changes in RNA sequences. Recent studies have reported that aberrant adenosine-to-inosine RNA editing is implicated in cancers. Until now, very few functionally important protein-recoding editing targets have been discovered. Here, we investigated the role of arecently discovered protein-recoding editing target COPA (coatomer subunit α) in hepatocellular carcinoma (HCC). Clinical implication of COPA editing was studied in a cohort of 125 HCC patients. CRISPR/Cas9-mediated knockout of the editing site complementary sequence (ECS) was used to delete edited COPA transcripts endogenously. COPA editing-mediated change in its transcript or protein stability was investigated upon actinomycin D or cycloheximide treatment, respectively. Functional difference in tumourigenesis between wild-type and edited COPA (COPAWTvs. COPAI164V) and the exact mechanisms were also studied in cell models and mice. ADAR2 binds to double-stranded RNA formed between edited exon 6 and the ECS at intron 6 of COPA pre-mRNA, causing an isoleucine-to-valine substitution at residue 164. Reduced editing of COPA is implicated in the pathogenesis of HCC, and more importantly, it may be involved in many cancer types. Upon editing, COPAWT switches from a tumour-promoting gene to a tumour suppressor that has a dominant-negative effect. Moreover, COPAI164V may undergo protein conformational change and therefore become less stable than COPAWT. Mechanistically, COPAI164V may deactivate the PI3K/AKT/mTOR pathway through downregulation of caveolin-1 (CAV1). We uncover an RNA editing-associated mechanism of hepatocarcinogenesis by which downregulation of ADAR2 caused the loss of tumour suppressive COPAI164V and concurrent accumulation of tumour-promoting COPAWT in tumours; a rapid degradation of COPAI164V protein and hyper-activation of the PI3K/AKT/mTOR pathway further promote tumourigenesis. RNA editing is a process in which RNA is changed after it is made from DNA, resulting in an altered gene product. In this study, we found that RNA editing of a gene known as coatomer subunit α (COPA) is lower in tumour samples and discovered that this editing process changes COPA protein from a tumour-promoting form to a tumour-suppressive form. Loss of the edited COPA promotes the development of liver cancer.

Bi-FoRe: an efficient bidirectional knockin strategy to generate pairwise conditional alleles with fluorescent indicators

Gene expression labeling and conditional manipulation of gene function are important for elaborate dissection of gene function. However, contemporary generation of pairwise dual-function knockin alleles to achieve both conditional and geno-tagging effects with a single donor has not been reported. Here we first developed a strategy based on a flipping donor named FoRe to generate conditional knockout alleles coupled with fluorescent allele-labeling through NHEJ-mediated unidirectional targeted insertion in zebrafish facilitated by the CRISPR/Cas system. We demonstrated the feasibility of this strategy at sox10 and isl1 loci, and successfully achieved Cre-induced conditional knockout of target gene function and simultaneous switch of the fluorescent reporter, allowing generation of genetic mosaics for lineage tracing. We then improved the donor design enabling efficient one-step bidirectional knockin to generate paired positive and negative conditional alleles, both tagged with two different fluorescent reporters. By introducing Cre recombinase, these alleles could be used to achieve both conditional knockout and conditional gene restoration in parallel; furthermore, differential fluorescent labeling of the positive and negative alleles enables simple, early and efficient real-time discrimination of individual live embryos bearing different genotypes prior to the emergence of morphologically visible phenotypes. We named our improved donor as Bi-FoRe and demonstrated its feasibility at the sox10 locus. Furthermore, we eliminated the undesirable bacterial backbone in the donor using minicircle DNA technology. Our system could easily be expanded for other applications or to other organisms, and coupling fluorescent labeling of gene expression and conditional manipulation of gene function will provide unique opportunities to fully reveal the power of emerging single-cell sequencing technologies.

Gene expression profiling of proximal and distal renal tubules in Atlantic salmon (Salmo salar) acclimated to fresh water and seawater.

Euryhaline teleost kidneys undergo a major functional switch from being filtratory in freshwater (FW) to being predominantly secretory in seawater (SW) conditions. The transition involves both vascular and tubular effects. There is consensus that the glomerular filtration rate is greatly reduced upon exposure to hyperosmotic conditions. Yet, regulation at the tubular level has only been examined sporadically in a few different species. This study aimed to obtain a broader understanding of transcriptional regulation in proximal versus distal tubular segments during osmotic transitions. Proximal and distal tubule cells were dissected separately by laser capture microdissection, RNA was extracted, and relative mRNA expression levels of >30 targets involved in solute and water transport were quantified by quantitative PCR in relation to segment type in fish acclimated to FW or SW. The gene categories were aquaporins, solute transporters, fxyd proteins, and tight junction proteins. aqp8bb1, aqp10b1, nhe3, sglt1, slc41a1, cnnm3, fxyd12a, cldn3b, cldn10b, cldn15a, and cldn12 were expressed at a higher level in proximal compared with distal tubules. aqp1aa, aqp1ab, nka-a1a, nka-a1b, nkcc1a, nkcc2, ncc, clc-k, slc26a6C, sglt2, fxyd2, cldn3a, and occln were expressed at a higher level in distal compared with proximal tubules. Expression of aqp1aa, aqp3a1, aqp10b1, ncc, nhe3, cftr, sglt1, slc41a1, fxyd12a, cldn3a, cldn3b, cldn3c, cldn10b, cldn10e, cldn28a, and cldn30c was higher in SW- than in FW-acclimated salmon, whereas the opposite was the case for aqp1ab, slc26a6C, and fxyd2. The data show distinct segmental distribution of transport genes and a significant regulation of tubular transcripts when kidney function is modulated during salinity transitions.

Blueberry Counteracts BV-2 Microglia Morphological and Functional Switch after LPS Challenge.

Microglia, the innate immune cells of the CNS, respond to brain injury by activating and modifying their morphology. Our study arises from the great interest that has been focused on blueberry (BB) for the antioxidant and pharmacological properties displayed by its components. We analyzed the influence of hydroalcoholic BB extract in resting or lipopolysaccharide (LPS)-stimulated microglia BV-2 cells. BB exerted a protective effect against LPS-induced cytotoxicity, as indicated by cell viability. BB was also able to influence the actin cytoskeleton organization, to recover the control phenotype after LPS insult, and also to reduce LPS-driven migration. We evaluated the activity of Rho and Rac1 GTPases, which regulate both actin cytoskeletal organization and migratory capacity. LPS caused an increase in Rac1 activity, which was counteracted by BB extract. Furthermore, we demonstrated that, in the presence of BB, mRNA expression of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α decreased, as did the immunofluorescence signal of iNOS, whereas that of Arg-1 was increased. Taken together, our results show that, during the inflammatory response, BB extract shifts the M1 polarization towards the M2 phenotype through an actin cytoskeletal rearrangement. Based on that, we might consider BB as a nutraceutical with anti-inflammatory activities.

Rheostat positions: A new classification of protein positions relevant to pharmacogenomics.

To achieve the full potential of pharmacogenomics, one must accurately predict the functional outcomes that arise from amino acid substitutions in proteins. Classically, researchers have focused on understanding the consequences of individual substitutions. However, literature surveys have shown that most substitutions were created at evolutionarily conserved positions. Awareness of this bias leads to a shift in perspective, from considering the outcomes of individual substitutions to understanding the roles of individual protein positions. Conserved positions tend to act as “toggle” switches, with most substitutions abolishing function. However, nonconserved positions have been found equally capable of affecting protein function. Indeed, many nonconserved positions act like functional dimmer switches (“rheostat” positions): this is revealed when multiple substitutions are made at a single position. Each substitution has a different functional outcome; the set of substitutions spans a range of outcomes. Finally, some nonconserved positions appear neutral, capable of accommodating all amino acid types without modifying function. This paper reviews the currently-known properties of rheostat positions, with examples shown for pyruvate kinase, organic anion transporting polypeptide 1B1, the beta-lactamase inhibitory protein, and angiotensin-converting enzyme 2. Outcomes observed for rheostat positions have implications for the rational design of drug analogs and allosteric drugs. Furthermore, this new framework—comprising three types of protein positions—provides a new approach to interpreting disease and population-based databases of amino acid changes. In conclusion, although a full understanding of substitution outcomes at rheostat positions poses a challenge, utilization of this new frame of reference will further advance the application of pharmacogenomics.

1675-P: Adipocyte Adaptive Immune Cell Function and Exhaustion Contribute to the Drop in Adipose Tissue (AT) Regulatory T-Cells (Tregs) in Human Obesity

Background: AT Tregs are decreased in obese mice. We have previously shown that preventing this decline improves insulin sensitivity, despite persistent obesity, underscoring a key role for AT Tregs in systemic metabolism. Therefore, we sought to examine AT Treg changes in human subjects and elucidate potential mechanisms responsible for these changes. Methods: We obtained visceral (VAT) and subcutaneous adipose tissue (SAT) at elective surgery in 54 obese and 18 lean subjects, and SAT via liposuction in 15 lean subjects before and after 2 weeks high fat diet (HFD). Results: Obese individuals had 67% less Treg abundance in VAT (p<0.001) and 79% in SAT (p<0.001); Tregs as %CD4+ cells correlated with insulin sensitivity and inversely with BMI. Subjects ingesting a HFD experienced a 50% drop in SAT Tregs (p<0.001), which correlated with decreased insulin sensitivity by clamp. To understand this drop, we next performed gene expression analyses on isolated adipocytes, AT T cells, and VAT Tregs. Obese adipocytes had profound increases in major histocompatibility class II (MHCII) genes, but decreases in fatty acid oxidation and mitochondrial genes, suggesting a functional switch from a primary metabolic to an adaptive immune cell. These changes were associated with decreased AT Treg abundance, and increased insulin resistance and BMI. T cells revealed increased Tbet, a proinflammatory CD4+ Th1 cell marker, and decreased FOXP3, a Treg marker, consistent with our demonstration that human adipocytes can present antigen to activate CD4+ Th1 cells. Obese vs. lean VAT Tregs also had a 3-fold (p<0.05) increase in the T cell exhaustion marker, program cell death protein-1 (PD-1), and a 75% (p<0.05) decrease in CD127, which is attenuated in exhausted T cells. Discussion: Thus, human adipocyte immune cell activity, combined with Treg exhaustion, contributes to the drop in AT Tregs and strongly affects metabolism. Disclosure D. Bradley: None. A.M. Blaszczak: None. A. Smith: None. A.D. Jalilvand: None. V.P. Wright: None. K. Wyne: Advisory Panel; Self; Novo Nordisk Inc. Research Support; Self; Sanofi. S. Noria: None. J. Joseph: None. J.Z. Liu: None. W. Hsueh: None. Funding American Diabetes Association (1-16-ICTS-049 to W.H.); National Institutes of Health (KL2TR001068, R01HL135622)

Photosynthetic inner antenna CP47 plays important roles in ephemeral plants in adapting to high light stress

Throughout 3.5 billion years of evolution, photosynthesis of land plants has developed a complicated antenna system to cope with the ever-changing environments. The antenna system of photosystem (PS) II includes the outer antennae and inner antennae. The inner antennae CP43 and CP47, located in the closest peripheral of PSII reaction center (RC), play important roles in facilitating excitation energy transport from the outer antennae to the PSII RC. Although PSII RC is the last station of energy transport, it is the inner antenna CP47, not the RC, which possesses the lowest energy level in PSII. Berteroa incana (B. incana), which is a vascular plant grown in the Gobi region, can sustain very high photosynthesis capacity under very high light conditions. It has been discovered that the thylakoid membrane of B. incana possesses a unique low fluorescence emission spectrum because the fluorescence emission of CP47 (695 nm) is the main fluorescence emission peak of PSII. In this paper, the thylakoid membrane, isolated from B. incana grown under a light condition of 100 μM photons m−2 s-1 and subjected to high light treatment (1600 μM photons m-2 s-1 for 1.5 h or 3 h) was employed for the research. It has been found that the high fluorescence emission of CP47 decreased remarkably upon exposure to the high light treatment. Further observation revealed that the drastic changes in the CP47 fluorescence emission were accompanied by a slight reduction in the amount of CP47 and an enhancement of the CP29-LHCII-CP24 assembly. It is proposed that CP47 enables the functional switch between the excitation energy transfer towards PSII RC, and the overexcitation quenching in the PSII core. Together with CP43, CP47 plays important roles in regulating excitation energy distribution in PSII core complexes.

Repression of the lysogenic PR promoter in bacteriophage TP901-1 through binding of a CI-MOR complex to a composite OM-OR operator

A functional genetic switch from the lactococcal bacteriophage TP901-1, deciding which of two divergently transcribing promoters becomes most active and allows this bi-stable decision to be inherited in future generations requires a DNA region of less than 1 kb. The fragment encodes two repressors, CI and MOR, transcribed from the PR and PL promoters respectively. CI can repress the transcription of the mor gene at three operator sites (OR, OL, and OD), leading to the immune state. Repression of the cI gene, leading to the lytic (anti-immune) state, requires interaction between CI and MOR by an unknown mechanism, but involving a CI:MOR complex. A consensus for putative MOR binding sites (OM sites), and a common topology of three OM sites adjacent to the OR motif was here identified in diverse phage switches that encode CI and MOR homologs, in a search for DNA sequences similar to the TP901-1 switch. The OR site and all putative OM sites are important for establishment of the anti-immune repression of PR, and a putative DNA binding motif in MOR is needed for establishment of the anti-immune state. Direct evidence for binding between CI and MOR is here shown by pull-down experiments, chemical crosslinking, and size exclusion chromatography. The results are consistent with two possible models for establishment of the anti-immune repression of cI expression at the PR promoter.

Resting state functional network switching rate is differently altered in bipolar disorder and major depressive disorder.

The clinical misdiagnosis ratio of bipolar disorder (BD) patients to major depressive disorder (MDD) patients is high. Recent findings hypothesize that the ability to flexibly recruit functional neural networks is differently altered in BD and MDD patients. This study aimed to explore distinct aberrance of network flexibility during dynamic networks configuration in BD and MDD patients. Resting state functional magnetic resonance imaging of 40 BD patients, 61 MDD patients, and 61 matched healthy controls were recruited. Dynamic functional connectivity matrices for each subject were constructed with a sliding window method. Then, network switching rate of each node was calculated and compared among the three groups. BD and MDD patients shared decreased network switching rate of regions including left precuneus, bilateral parahippocampal gyrus, and bilateral dorsal medial prefrontal cortex. Apart from these regions, MDD patients presented specially decreased network switching rate in the bilateral anterior insula, left amygdala, and left striatum. Taken together, BD and MDD patients shared decreased network switching rate of key hubs in default mode network and MDD patients presented specially decreased switching rate in salience network and striatum. We found shared and distinct aberrance of network flexibility which revealed altered adaptive functions during dynamic networks configuration of BD and MDD.