A global transcriptional activator involved in the iron homeostasis in cyanobacteria.

Cyanobacteria use a series of adaptation strategies and a complicated regulatory network to maintain intracellular iron (Fe) homeostasis. Here, a global activator named IutR has been identified through three-dimensional chromosome organization and transcriptome analysis in a model cyanobacterium Synechocystis sp. PCC 6803. Inactivation of all three homologous IutR-encoding genes resulted in an impaired tolerance of Synechocystis to Fe deficiency and loss of the responses of Fe uptake-related genes to Fe-deplete conditions. Protein-promoter interaction assays confirmed the direct binding of IutR with the promoters of genes related to Fe uptake, and chromatin immunoprecipitation sequencing analysis further revealed that in addition to Fe uptake, IutR could regulate many other physiological processes involved in intracellular Fe homeostasis. These results proved that IutR is an important transcriptional activator, which is essential for cyanobacteria to induce Fe-deficiency response genes. This study provides in-depth insights into the complicated Fe-deficient signaling network and the molecular mechanism of cyanobacteria adaptation to Fe-deficient environments.

Targeting Magnaporthe oryzae effector MoErs1 and host papain-like protease OsRD21 interaction to combat rice blast.

Effector proteins secreted by plant pathogenic fungi are important artilleries against host immunity, but there is no precedent of such effectors being explored as antifungal targets. Here we demonstrate that MoErs1, a species-specific effector protein secreted by the rice blast fungus Magnaporthe oryzae, inhibits the function of rice papain-like cysteine protease OsRD21 involved in rice immunity. Disrupting MoErs1-OsRD21 interaction effectively controls rice blast. In addition, we show that FY21001, a structure-function-based designer compound, specifically binds to and inhibits MoErs1 function. FY21001 significantly and effectively controls rice blast in field tests. Our study revealed a novel concept of targeting pathogen-specific effector proteins to prevent and manage crop diseases.

Single-cell transcriptomics of the immune system in ME/CFS at baseline and following symptom provocation.

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a serious and poorly understood disease. To understand immune dysregulation in ME/CFS, we use single-cell RNA sequencing (scRNA-seq) to examine immune cells in patient and control cohorts. Postexertional malaise (PEM), an exacerbation of symptoms following strenuous exercise, is a characteristic symptom of ME/CFS. To detect changes coincident with PEM, we applied scRNA-seq on the same cohorts following exercise. At baseline, ME/CFS patients display classical monocyte dysregulation suggestive of inappropriate differentiation and migration to tissue. We identify both diseased and more normal monocytes within patients, and the fraction of diseased cells correlates with disease severity. Comparing the transcriptome at baseline and postexercise challenge, we discover patterns indicative of improper platelet activation in patients, with minimal changes elsewhere in the immune system. Taken together, these data identify immunological defects present at baseline in patients and an additional layer of dysregulation in platelets.

Molecular Basis and Genome Editing Applications of a Compact Eubacterium ventriosum CRISPR-Cas9 System.

CRISPR-Cas9 systems have been widely harnessed for diverse genome editing applications because of their ease of use and high efficiency. However, the large molecular sizes and strict PAM requirements of commonly used CRISPR-Cas9 systems restrict their broad applications in therapeutics. Here, we report the molecular basis and genome editing applications of a novel compact type II-A Eubacterium ventriosum CRISPR-Cas9 system (EvCas9) with 1107 residues and distinct 5'-NNGDGN-3' (where D represents A, T, or G) PAM specificity. We determine the cryo-EM structure of EvCas9 in a complex with an sgRNA and a target DNA, revealing the detailed PAM recognition and sgRNA and target DNA association mechanisms. Additionally, we demonstrate the robust genome editing capacity of EvCas9 in bacteria and human cells with superior fidelity compared to SaCas9 and SpCas9, and we engineer it to be efficient base editors by fusing a cytidine or adenosine deaminase. Collectively, our results facilitate further understanding of CRISPR-Cas9 working mechanisms and expand the compact CRISPR-Cas9 toolbox.

Identification of plant cadmium resistance gene family in Brassica napus and functional analysis of BnPCR10.1 involved in cadmium and copper tolerance.

The plant cadmium resistance (PCR) family proteins play important roles in maintaining metal homeostasis and detoxification. However, few functional PCR genes have been well-characterized in plants. In this study, we identified and cloned 26 BnPCR genes from the rapeseed (Brassica napus) genome. They were divided into four groups (I-IV) based on their phylogenetic relationship. Yeast functional complementation experiments showed that BnPCRs can transport copper (Cu) and cadmium (Cd) in yeast. The expression levels of the BnPCRs were variable among different organs. Moreover, most of the genes were induced by Cu2+ and Cd2+ stress. Among these genes, BnPCR10.1 was highly expressed in various organs and induced by Cu2+ and Cd2+. Therefore, we studied the function of BnPCR10.1 in more detail. BnPCR10.1 was localized to the plasma membrane (PM), and expression in yeast enhanced yeast cells to export Cu and Cd. Furthermore, overexpression of BnPCR10.1 transgenic lines pro35S::BnPCR10.1;athma5 had lower concentration of Cu in roots than athma5 mutants. In addition, transgenic plants pro35S::BnPCR10.1;atpdr8 had lower concentration of Cd in shoots and roots than atpdr8 mutants. Net Cu2+ and Cd2+ efflux assay showed that there was decreased absorption of Cu2+ and Cd2+ in the transgenic Arabidopsis elongation zone of roots than in athma5 and atpdr8 mutants, respectively. These results provide new information on BnPCRs and their roles in response to heavy metals and reveal the mechanism used by BnPCR10.1 to detoxify Cu and Cd. Our findings facilitate a theoretical basis for the genetic improvement of Cu-Cd tolerance in rapeseed.

m6 A-Dependent Modulation via IGF2BP3/MCM5/Notch Axis Promotes Partial EMT and LUAD Metastasis.

The importance of mRNA N6-methyladenosine (m6 A) modification during tumor metastasis is controversial as it plays distinct roles in different biological contexts. Moreover, how cancer cell plasticity is shaped by m6 A modification is interesting but remains uncharacterized. Here, this work shows that m6 A reader insulin like growth factor 2 mRNA binding protein 3 (IGF2BP3) is remarkably upregulated in metastatic lung adenocarcinoma (LUAD) and indicates worse prognosis of patients. Interestingly, IGF2BP3 induces partial epithelial-mesenchymal-transition (EMT) and confers LUAD cells plasticity to metastasize through m6 A-dependent overactivation of Notch signaling. Mechanistically, IGF2BP3 recognized m6 A-modified minichromosome maintenance complex component (MCM5) mRNAs to prolong stability of them, subsequently upregulating MCM5 protein, which competitively inhibits SIRT1-mediated deacetylation of Notch1 intracellular domain (NICD1), stabilizes NICD1 protein and contributes to m6 A-dependent IGF2BP3-mediated cellular plasticity. Notably, a tight correlation of the IGF2BP3/MCM5/Notch axis is evidenced in clinical LUAD specimens. Therefore, this study elucidates a critical role of m6 A modification on LUAD cell plasticity in fostering tumor metastasis via the above axis, providing potential targets for metastatic LUAD.

Ethnicity Disparities in the Prevalence, Awareness, Treatment, and Control Rates of Hypertension in China.

Objectives: Previous studies reported that there were disparities in hypertension management among different ethnic groups, and this study aimed to systematically determine the prevalence, awareness, treatment, and control rates of hypertension in multiple Chinese ethnic groups. Methods: We searched Embase, PubMed, and Web of Science for articles up to 25 October, 2022. The pooled prevalence, awareness, treatment, and control rates of hypertension were estimated with 95% confidence intervals (CI). The heterogeneity of estimates among studies was assessed by the Cochran Q test and I 2 statistic. Meta-regression analyses were conducted to identify the factors influencing the heterogeneity of the pooled prevalence, awareness, treatment, and control rate of hypertension. Results: In total, 45 publications including 193,788 cases and 587,826 subjects were eligible for the analyses. The lowest prevalence was found in the Han group (27.0%), and the highest prevalence was in the Mongolian population (39.8%). The awareness rates ranged from 24.4% to 58.0% in the four ethnic groups. Both the highest treatment and control rates were found in the Mongolian population (50.6% and 16.0%, respectively), whereas the Yi group had the lowest control rate (8.0%). In addition, the study year, the mean age of subjects, mean body mass index of subjects, tobacco use (%), alcohol use (%), residence (urban%), and education (primary school%) had varied effects on heterogeneity. Conclusions: These findings highlight the disparities in prevalence, awareness, treatment, and control rates of hypertension in a different ethnic population of China, which could provide suggestions for making targeted prevention measures.

The association between altitude and the prevalence of hypertension among permanent highlanders.

Hypertension (HTN) is a growing contributor to the global disease burden, and it is prevalent among people living at high altitudes (H-ALTs). This study aimed to explore the relationship between altitude and the prevalence of HTN among inhabitants living at H-ALTs. We searched electronic databases, including PubMed, Embase, and Web of Science, up to April 30, 2022. The quality of included studies was assessed using the Joanna Briggs Institute (JBI) checklist for prevalence studies. A total of 1273 articles were screened, and 32 studies (86,487 participants) were eligible for further analyses. The pooled prevalence among highlanders was 28.7%. General additive model (GAM)-based meta-regression analysis was conducted to explore the association between altitude and the prevalence of HTN. A curve-shaped line was found between altitude and the prevalence of HTN (ß = 0.998, p = 0.039) after adjusting for factors including publication year, sample size, age, sex, ethnic group, body mass index (BMI), smoking and alcohol consumption. The turning point was observed at 3300 m. The predictive parameter indicated that the smoothness and goodness of model fit were good (GCV = 0.014, R2 = 0.60, respectively). The findings may provide clues for further mechanistic studies that can improve HTN prevention among highlanders.

Metastatic Tumor Cell-Specific FABP7 Promotes NSCLC Metastasis via Inhibiting ß-Catenin Degradation.

Metastasis accounts for 90% of cancer-related deaths and represents a prominent malignant feature in non-small cell lung cancer (NSCLC), while tumor cell-specific mechanisms and molecules pivotal for the metastatic capacity remain unclear. By analyzing single-cell RNA sequencing data, we found that fatty acid binding protein 7 (FABP7) was specifically up-regulated in tumor cells of metastatic NSCLC patients and might be a prognostic indicator for poor survival. Experimental studies based on NSCLC cell lines showed that FABP7 promoted the metastatic competencies of NSCLC cells in vitro and in vivo. Mechanistically, we demonstrated that FABP7 was important to canonical Wnt signaling activation and competitively inhibited the interaction between ß-catenin and components of its cytoplasmic degradation complex, thereby repressing the phosphorylation-dependent ubiquitination and degradation of ß-catenin. Our present study identifies FABP7 as a metastatic tumor cell-specific pro-metastatic gene and uncovers a previously unknown regulatory mechanism underlying Wnt hyperactivation via FABP7-impaired cytoplasmic ß-catenin degradation, implicating a novel molecule in regulating NSCLC metastasis.

Programmed genome editing by a miniature CRISPR-Cas12f nuclease.

The RNA-guided CRISPR-associated (Cas) nucleases are versatile tools for genome editing in various organisms. The large sizes of the commonly used Cas9 and Cas12a nucleases restrict their flexibility in therapeutic applications that use the cargo-size-limited adeno-associated virus delivery vehicle. More compact systems would thus offer more therapeutic options and functionality for this field. Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5' T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA. We show that AsCas12f1 functions as an effective genome-editing tool in both bacteria and human cells using various delivery methods, including plasmid, ribonucleoprotein and adeno-associated virus. The small size of AsCas12f1 offers advantages for cellular delivery, and characterizations of AsCas12f1 may facilitate engineering more compact genome-manipulation technologies.

Electronic characteristics of BRCA1 mutations in DNA.

The efficient and low-cost way for gene mutation detection and identification are conducive for the detection of disease. Here, we report the electronic characteristics of the gene of breast cancer 1 in four common mutation types: duplication, single nucleotide variant, deletion, and indel. The electronic characteristics are investigated by the combination of density functional theory and non-equilibrium Green's function formulation with decoherence. The magnitude of conductance of these DNA molecules and mutational changes are found to be detectable experimentally. In this study, we also find the significant mutation type dependent on the change of conductance. Hence these mutations are expected to be identifiable. We find deletion type mutation shows the largest change in relative conductance (~97%), whereas the indel mutation shows the smallest change in relative conductance (~27%). Therefore, this work presents a possibility of electronic detection and identification of mutations in DNA, which could be an efficient method as compared to the conventional methods.

Elevated cadmium and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels in residents living near electroplating industries.

This study aims to evaluate the body burdens of cadmium (Cd) associated with potential health impairment in residents living near electroplating industries. A total of 269 residents from exposure area and 106 from control area were recruited. We measured the blood and urinary Cd levels using an inductively coupled plasma mass spectrometer (ICP-MS); performed physical examinations; determined the urinary levels of ß2-microglobulin (ß2-MG), Nacetyl-ß-D-glucosaminidase (NAG), and 8-hydroxy-2'-deoxyguanosine (8-OHdG); and evaluated the associations between Cd and these biomarkers. Blood and urinary Cd levels in exposure group were statistically higher than in control group (1.712 vs. 1.159 µg/L; 1.980 vs. 1.740 µg/L, respectively, p < 0.05). Urinary ß2-MG and 8-OHdG levels in exposure group were also statistically higher (0.448 vs. 0.090 mg/L; 12.759 vs. 12.115 µg/L, respectively, p < 0.05), but urinary NAG levels showed no significant difference between the two groups (13.614 vs. 8.246 IU/L, p > 0.05). The proportion of abnormal nasal symptoms occurring in exposed subjects (88.8%) was much higher than in control subjects (78.2%, p < 0.05). Urinary Cd levels were positively correlated with blood Cd levels, urinary 8-OHdG, and NAG levels (r = 0.307, r = 0.185, r = 0.150, p < 0.05), but not correlated with urinary ß2-MG levels (p > 0.05). In conclusion, our study revealed that residents living in close proximity to electroplating industries had elevated body burdens of Cd levels, as well as slight renal dysfunction and DNA oxidation damage.

Genetic diversity and natural selection on the thrombospondin-related adhesive protein (TRAP) gene of Plasmodium falciparum on Bioko Island, Equatorial Guinea and global comparative analysis.

BACKGROUND: Thrombospondin-related adhesive protein (TRAP) is a transmembrane protein that plays a crucial role during the invasion of Plasmodium falciparum into liver cells. As a potential malaria vaccine candidate, the genetic diversity and natural selection of PfTRAP was assessed and the global PfTRAP polymorphism pattern was described. METHODS: 153 blood spot samples from Bioko malaria patients were collected during 2016-2018 and the target TRAP gene was amplified. Together with the sequences from database, nucleotide diversity and natural selection analysis, and the structural prediction were preformed using bioinformatical tools. RESULTS: A total of 119 Bioko PfTRAP sequences were amplified successfully. On Bioko Island, PfTRAP shows its high degree of genetic diversity and heterogeneity, with π value for 0.01046 and Hd for 0.99. The value of dN-dS (6.2231, p < 0.05) hinted at natural selection of PfTRAP on Bioko Island. Globally, the African PfTRAPs showed more diverse than the Asian ones, and significant genetic differentiation was discovered by the fixation index between African and Asian countries (Fst > 0.15, p < 0.05). 667 Asian isolates clustered in 136 haplotypes and 739 African isolates clustered in 528 haplotypes by network analysis. The mutations I116T, L221I, Y128F, G228V and P299S were predicted as probably damaging by PolyPhen online service, while mutations L49V, R285G, R285S, P299S and K421N would lead to a significant increase of free energy difference (ΔΔG > 1) indicated a destabilization of protein structure. CONCLUSIONS: Evidences in the present investigation supported that PfTRAP gene from Bioko Island and other malaria endemic countries is highly polymorphic (especially at T cell epitopes), which provided the genetic information background for developing an PfTRAP-based universal effective vaccine. Moreover, some mutations have been shown to be detrimental to the protein structure or function and deserve further study and continuous monitoring.

Personal exposure to PM2.5-bound heavy metals associated with cardiopulmonary function in general population.

To better understand the cardiopulmonary alterations associated with personal exposed PM2.5-bound heavy meals, we conducted a cross-sectional study in 2018 on 54 general residents. For each subject, PM2.5 exposure filter was collected by a low-volume sampler for 24 h; blood and urine samples were collected subsequently. Heavy metals in PM2.5, blood, and urine samples were determined by inductively coupled plasma mass spectrometry method. PM2.5-bound Mn, Cd, Sb, Pb, and Ni levels were 20.5, 9.27, 9.59, 28.3, and 16.9 ng/m3, respectively. The distribution of these metals followed the order: Pb (33.47%) > Mn (24.24%) > Ni (19.99%) > Sb (11.34%) > Cd (10.96%). The distribution of heavy meals in PM2.5, blood, and urine differed from each other. PM2.5-bound Cd, Pb levels were positively correlated with blood Cd, Pb levels (r = 0.323, r = 0.334, p < 0.05), respectively. PM2.5-bound Cd level was significantly higher in smoking group than non-smoking group (28.8 vs. 7.27 ng/m3, p < 0.01), same as Sb level (12.0 vs. 9.34 ng/m3, p < 0.01). Cd and Pb exposure might interact with cardiovascular function through autonomic regulation. No significant correlation was observed between metal exposure and pulmonary function. In conclusion, our data suggested that personal exposure to specific PM2.5-bound heavy metals might interact with profound cardiovascular alterations.

The prevalence of smoking in male medical students from 16 Chinese medical colleges and the associated factors.

BACKGROUND: The prevalence of smoking among Chinese medical students in medical colleges is a matter of significant concern. This study aimed to explore the prevalence of smoking and the associated factors among these students. METHODS: A total of 7,728 students from 16 colleges in 12 provinces were surveyed to measure their smoking behaviors, attitudes toward tobacco control, and knowledge about smoking risk. An evaluation of campus environment and policy was conducted at the same time. Frequencies were calculated using weighted estimators for complex sampling. A two-level logistic regression model was used to assess factors associated with current smoking. RESULTS: The prevalence of smoking among male medical students in the 16 medical colleges was 12.8%. Students' participation in smoking, smoking initiation after school enrollment, and intention-to-quit-smoking rates may be influenced by their anti-tobacco attitude, the belief that smoking can help to relieve stress, their parents' smoking behaviors, their best friend's attitude towards smoking and second-hand smoke exposure (SHSE), and their awareness of tobacco hazards. CONCLUSIONS: This study provides sufficient evidence to support the implementation of systematic intervention strategies to decrease the smoking rate among male medical students. These strategies may include tobacco control education, smoking cessation technologies, communication, and the enforcement of smoke-free policies to create a tobacco-controlled campus environment.

A Global View on Prevalence of Hypertension and Human Develop Index.

Background: The incidence of hypertension has been increasing in the past decade. Little is known regarding the relationship between hypertension and human development index (HDI). Objectives: The objective is to identify the relationship between the prevalence of hypertension and human development index (HDI). Methods: An ecological study was conducted. The data from World Health Organization reports and United Nations Development Programme reports for 182 countries, including the HDI values, rates of tobacco use, physical inactivity, alcohol use, and salt intake. The Generalized Additive Models were implemented to assess the association between the prevalence of hypertension and the HDI. Results: Among 182 countries, the prevalence of hypertension ranged from 13% to 41%. The highest HDI value was 0.949 and the lowest was 0.352. In model 1, statistically significant associations were found in three populations, the largest R2 was 0.245. In model 2, the largest R2 was 0.485. In linear part, there was negative relationship in female population, while HDI was associated with prevalence of hypertension in all three populations, which was explained by spline function. The curve indicated that there were three intervals from low to high HDI. From 0 to 0.6 and 0.8 to 1, an evident decreasing trend of prevalence was found, while the rate increased when HDI was in the interval of 0.6 to 0.8. Conclusions: In this study, we identified the association between the prevalence of hypertension and the HDI and the underlying pattern of the relationship. The findings will aid the planning of hypertension control priorities and provide suggestions for interventions.

Metabolism and Toxicity of Emodin: Genome-Wide Association Studies Reveal Hepatocyte Nuclear Factor 4α Regulates UGT2B7 and Emodin Glucuronidation.

Emodin is the main toxic component in Chinese medicinal herbs such as rhubarb. Our previous studies demonstrated that genetic polymorphisms of UDP-glucuronosyltransferase 2B7 (UGT2B7) had an effect on the glucuronidation and detoxification of emodin. This study aimed to reveal the transcriptional regulation mechanism of UGT2B7 on emodin glucuronidation and its effect on toxicity. Emodin glucuronic activity and genome and transcriptome data were obtained from 36 clinical human kidney tissues. The genome-wide association studies (GWAS) identified that four single nucleotide polymorphisms (SNPs) (rs6093966, rs2868094, rs2071197, and rs6073433), which were located on the hepatocyte nuclear factor 4α (HNF4A) gene, were significantly associated with the emodin glucuronidation (p < 0.05). Notably, rs2071197 was significantly associated with the gene expression of HNF4A and UGT2B7 and the glucuronidation of emodin. The gene expression of HNF4A showed a high correlation with UGT2B7 (R2 = 0.721, p = 5.83 × 10-11). The luciferase activity was increased 7.68-fold in 293T cells and 2.03-fold in HepG2 cells, confirming a significant transcriptional activation of UGT2B7 promoter by HNF4A. The knockdown of HNF4A in HepG2 cells (36.6%) led to a significant decrease of UGT2B7 (19.8%) and higher cytotoxicity (p < 0.05). The overexpression of HNF4A in HepG2 cells (31.2%) led to a significant increase of UGT2B7 (24.4%) and improved cell viability (p < 0.05). Besides, HNF4A and UGT2B7 were both decreased in HepG2 cells and rats after treatment with emodin. In conclusion, emodin used long term or in high doses could inhibit the expression of HNF4A, thereby reducing the expression of UGT2B7 and causing hepatotoxicity.

A follow-up study on the characterization and health risk assessment of heavy metals in ambient air particles emitted from a municipal waste incinerator in Zhejiang, China.

To confirm our hypothesis that inhalation might be the primary exposure route of heavy metals for children living in proximity to a municipal waste incinerator (MWI), we conducted a one-year follow up study to characterize the distributions of heavy metals featured in different types of ambient air particles, including PM1, PM2.5 and PM10, at two exposure sites near the MWI (E1 and E2) and one control site (C) in Zhejiang, China. Particle samples were collected by a mid-volume sampler and heavy metals were determined by the inductively coupled plasma mass spectrometry method. The mass concentrations of PM1, PM2.5 and PM10 were 52.0, 85.8 and 100.3 µg/m3 at E1 site, while the concentrations were 40.2, 92.1 and 106.6 µg/m3 at E2 site and 33.4, 55.6 and 66.1 µg/m3 at C site, respectively. Both E1 and E2 had higher PM1, PM2.5 and PM10 levels than C site. The levels of pollution were season dependent, with autumn having the highest levels of PM1, PM2.5 and PM10 across all three sampling sites. Regarding the distributions of heavy metals, Pb accounted for the majority of the seven metals in all groups, ranging from 43.2% to 51.3%, followed by Mn that ranged from 22.0% to 32.0%. The Pb levels of PM1, PM2.5 and PM10 in the MWI area were 22.6, 34.2 and 36.2 ng/m3, respectively, while Mn levels were 10.1, 20.0 and 23.5 ng/m3, respectively. The health risk assessment results suggested that residents were suffering high non-carcinogenic risk posed by MWI-emitted particle-bound toxic metals, as well as the high lifetime carcinogenic risk. This study revealed that ambient air, no matter whether near or far away from an MWI, bore more PM1, PM2.5 and PM10 particles than general, non-polluted ambient air, especially in autumn.

Selective cadmium regulation mediated by a cooperative binding mechanism in CadR.

Detoxification of the highly toxic cadmium element is essential for the survival of living organisms. Pseudomonas putida CadR, a MerR family transcriptional regulator, has been reported to exhibit an ultraspecific response to the cadmium ion. Our crystallographic and spectroscopic studies reveal that the extra cadmium selectivity of CadR is mediated by the unexpected cooperation of thiolate-rich site I and histidine-rich site II. Cadmium binding in site I mediates the reorientation of protein domains and facilitates the assembly of site II. Subsequently, site II bridge-links 2 DNA binding domains through ligands His140/His145 in the C-terminal histidine-rich tail. With dynamic transit between 2 conformational states, this bridge could stabilize the regulator into an optimal conformation that is critical for enhancing the transcriptional activity of the cadmium detoxification system. Our results provide dynamic insight into how nature utilizes the unique cooperative binding mechanism in multisite proteins to recognize cadmium ions specifically.