UFGI publication round-up week 5/7
Determination of the optimum contribution of Brahman genetics in an Angus-Brahman multibreed herd for regulation of body temperature during hot weather.
Author information
- 1
- Department of Animal Science, Faculty of Veterinary Medicine, Uludag University, Bursa, Turkey.
- 2
- Department of Animal Sciences, University of Florida, Gainesville, FL.
Abstract
The objective was to evaluate the influence of varying amounts of Brahman genetics on body temperature under pasture conditions during hot weather. Vaginal temperatures were measured at 5-min intervals for 3 to 5 d on four occasions during August and September from a total of 190 pregnant cows that were either Angus, 2/8 Brahman (remainder Angus), Brangus (3/8 Brahman), 4/8 Brahman, 6/8 Brahman or Brahman. Vaginal temperature was higher for the first two replicates than for the second two replicates. In the first two replicates, average vaginal temperature did not differ between genetic groups, but average vaginal temperature from 1500 to 1900 h was lower for Brahman than other groups. In the second two replicates, average vaginal temperature was lower for cows that were 4/8 or higher Brahman than for cows that were 2/8 Brahman or Angus. Average vaginal temperature from 1500 to 1900 h was lower for cows that were 4/8 or higher Brahman than for cows that were 2/8 Brahman or Angus. In addition, Brahman cows had lower vaginal temperatures than cows that were 4/8 Brahman or 3/8 Brahman (i.e., Brangus). In one replicate, a tracking device was used to map cow location. At 1200 to 1300 h, cows that were 6/8 Brahman or Brahman had fewer observations near the tree line (i.e., in shade) than cows that were 4/8 Brahman or less. At 1500 to 1600 h, cows that were 4/8 or higher Brahman experienced fewer observations near the tree line than cows that contained a lower fraction of Brahman genetics. In summary, a minimum of 4/8 Brahman genetics was required to increase the ability to regulate body temperature and at least 6/8 Brahman when heat stress was severe. It is likely, therefore, that using Brahman genetics to optimize adaptation to thermal stress under conditions of severe heat stress requires a preponderance of Brahman genes.
Comparison of the Chinese bamboo partridge and red Junglefowl genome sequences highlights the importance of demography in genome evolution.
Author information
- 1
- Department of Biology, University of Florida, Gainesville, FL, 32611, USA. george.tiley@duke.edu.
- 2
- Department of Biology, Duke University, Durham, NC, 27708, USA. george.tiley@duke.edu.
- 3
- Department of Biology, University of Florida, Gainesville, FL, 32611, USA.
Abstract
BACKGROUND:
Recent large-scale whole genome sequencing efforts in birds have elucidated broad patterns of avian phylogeny and genome evolution. However, despite the great interest in economically important phasianids like Gallus gallus (Red Junglefowl, the progenitor of the chicken), we know little about the genomes of closely related species. Gallus gallus is highly sexually dichromatic and polygynous, but its sister genus, Bambusicola, is smaller, sexually monomorphic, and monogamous with biparental care. We sequenced the genome of Bambusicola thoracicus (Chinese Bamboo Partridge) using a single insert library to test hypotheses about genome evolution in galliforms. Selection acting at the phenotypic level could result in more evidence of positive selection in the Gallus genome than in Bambusicola. However, the historical range size of Bambusicola was likely smaller than Gallus, and demographic effects could lead to higher rates of nonsynonymous substitution in Bambusicola than in Gallus.
RESULTS:
We generated a genome assembly suitable for evolutionary analyses. We examined the impact of selection on coding regions by examining shifts in the average nonsynonymous to synonymous rate ratio (dN/dS) and the proportion of sites subject to episodic positive selection. We observed elevated dN/dS in Bambusicola relative to Gallus, which is consistent with our hypothesis that demographic effects may be important drivers of genome evolution in Bambusicola. We also demonstrated that alignment error can greatly inflate estimates of the number of genes that experienced episodic positive selection and heterogeneity in dN/dS. However, overall patterns of molecular evolution were robust to alignment uncertainty. Bambusicola thoracicus has higher estimates of heterozygosity than Gallus gallus, possibly due to migration events over the past 100,000 years.
CONCLUSIONS:
Our results emphasized the importance of demographic processes in generating the patterns of variation between Bambusicola and Gallus. We also demonstrated that genome assemblies generated using a single library can provide valuable insights into avian evolutionary history and found that it is important to account for alignment uncertainty in evolutionary inferences from draft genomes.
Dicer cleaves 5′-extended microRNA precursors originating from RNA polymerase II transcription start sites.
Author information
- 1
- Department of Biochemistry and Molecular Biology.
- 2
- UF Health Cancer Center.
- 3
- Department of Microbiology & Cell Science, Institute of Food and Agricultural Sciences.
- 4
- Interdisciplinary Center for Biotechnology Research.
- 5
- UF Genetics Institute, University of Florida, Gainesville, FL 32610, USA.
Abstract
MicroRNAs (miRNAs) are approximately 22 nucleotide (nt) long and play important roles in post-transcriptional regulation in both plants and animals. In animals, precursor (pre-) miRNAs are ∼70 nt hairpins produced by Drosha cleavage of long primary (pri-) miRNAs in the nucleus. Exportin-5 (XPO5) transports pre-miRNAs into the cytoplasm for Dicer processing. Alternatively, pre-miRNAs containing a 5′ 7-methylguanine (m7G-) cap can be generated independently of Drosha and XPO5. Here we identify a class of m7G-capped pre-miRNAs with 5′ extensions up to 39 nt long. The 5′-extended pre-miRNAs are transported by Exportin-1 (XPO1). Unexpectedly, a long 5′ extension does not block Dicer processing. Rather, Dicer directly cleaves 5′-extended pre-miRNAs by recognizing its 3′ end to produce mature 3p miRNA and extended 5p miRNA both in vivo and in vitro. The recognition of 5′-extended pre-miRNAs by the Dicer Platform-PAZ-Connector (PPC) domain can be traced back to ancestral animal Dicers, suggesting that this previously unrecognized Dicer reaction mode is evolutionarily conserved. Our work reveals additional genetic sources for small regulatory RNAs and substantiates Dicer’s essential role in RNAi-based gene regulation.
Longitudinal timed function tests in Duchenne muscular dystrophy: ImagingDMD cohort natural history.
Author information
- 1
- Department of Physical Therapy, University of Florida, Gainesville, Florida, USA.
- 2
- The Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
- 3
- Oregon Health & Science University, Portland, Oregon, USA.
- 4
- Department of Statistics, University of Florida, Gainesville, Florida, USA.
- 5
- Department of Statistics & Data Sciences, The University of Texas at Austin, Texas, USA.
- 6
- Nemours Children’s Hospital, Orlando, Florida, USA.
- 7
- Department of Pediatrics and Molecular Genetics and Microbiology, Powell Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
- 8
- Department of Physiology and Functional Genomics, University of Florida, Gainesville, Florida, USA.
- 9
- Department of Pharmacology and Therapeutics, University of Florida, Gainesville, Florida, USA.
Abstract
INTRODUCTION:
Tests of ambulatory function are common clinical trial endpoints in Duchenne muscular dystrophy (DMD). The ImagingDMD study has generated a large data set using these tests, which can describe the contemporary natural history of DMD in 5-12.9 year olds.
METHODS:
92 corticosteroid treated boys with DMD and 45 controls participated in this longitudinal study. Subjects performed the 6 minute walk test (6MWT) and timed function tests (TFTs: 10m walk/run, 4 stairs, supine to stand).
RESULTS:
Boys with DMD had impaired functional performance even at 5-6.9 years. Boys older than 7 had significant declines in function over 1 year for 10m walk/run and 6MWT. 80% of subjects could perform all functional tests at 9 years old. TFTs appear to be slightly more responsive and predictive of disease progression than 6MWT in 7-12.9 year olds.
DISCUSSION:
This study provides insight into the contemporary natural history of key functional endpoints in DMD. This article is protected by copyright. All rights reserved.
Regulatory protein SrpA controls phage infection and core cellular processes in Pseudomonas aeruginosa.
Author information
- 1
- State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology of the Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China.
- 2
- Department of Microbiology, State Key Laboratory of Medicinal Chemical Biology, Key Laboratory of Molecular Microbiology and Technology of the Ministry of Education, College of Life Sciences, Nankai University, Tianjin, 300071, China.
- 3
- Department of Molecular Genetics and Microbiology, College of Medicine, University of Florida, Gainesville, FL, 32610, USA.
- 4
- The Key Laboratory of Industrial Biotechnology, Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi, 214122, China. raozhm@jiangnan.edu.cn.
- 5
- Department of Microbiology, State Key Laboratory of Medicinal Chemical Biology, Key Laboratory of Molecular Microbiology and Technology of the Ministry of Education, College of Life Sciences, Nankai University, Tianjin, 300071, China. wuweihui@nankai.edu.cn.
- 6
- State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology of the Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China. hongjiangyang@tust.edu.cn.
Abstract
Our understanding of the molecular mechanisms behind bacteria-phage interactions remains limited. Here we report that a small protein, SrpA, controls core cellular processes in response to phage infection and environmental signals in Pseudomonas aeruginosa. We show that SrpA is essential for efficient genome replication of phage K5, and controls transcription by binding to a palindromic sequence upstream of the phage RNA polymerase gene. We identify potential SrpA-binding sites in 66 promoter regions across the P. aeruginosa genome, and experimentally validate direct binding of SrpA to some of these sites. Using transcriptomics and further experiments, we show that SrpA, directly or indirectly, regulates many cellular processes including cell motility, chemotaxis, biofilm formation, pyocyanin synthesis and protein secretion, as well as virulence in a Caenorhabditis elegans model of infection. Further research on SrpA and similar proteins, which are widely present in many other bacteria, is warranted.
Proton pump inhibitors: from CYP2C19 pharmacogenetics to precision medicine.
Author information
- 1
- a Department of Pharmacotherapy and Translational Research, College of Pharmacy , University of Florida , Gainesville , FL , USA.
- 2
- b Center for Pharmacogenomics and Translational Research , Nemours, Children’s Health System , Jacksonville , FL , USA.
Abstract
Proton Pump inhibitors (PPIs) are commonly used for a variety of acid related disorders. Despite the overall effectiveness and safety profile of PPIs, some patients do not respond adequately or develop treatment related adverse events. This variable response among patients is in part due to genotype variability of CYP2C19, the gene encoding the CYP450 (CYP2C19) isoenzyme responsible for PPIs metabolism. Areas covered: This article provides an overview of the pharmacokinetics and mechanism of action of the currently available PPIs, including the magnitude of CYPC19 contribution to their metabolism. Additionally, the role of CYP2C19 genetic variability in the therapeutic effectiveness or outcomes of PPI therapy is highlighted in details, to provide supporting evidence for the potential value of CYP2C19 genotype-guided approaches to PPI drug therapy. Expert opinion: There is a large body of evidence describing the impact of CYP2C19 variability on PPIs and its potential role in individualizing PPI therapy, yet, CYP2C19 pharmacogenetics has not been widely implemented into clinical practice. More data are needed but CYP2C19 genotype-guided dosing of PPIs is likely to become increasingly common and is expected to improve clinical outcomes, and minimize side effects related to PPIs.
A new “American” subgroup of African-lineage Chikungunya virus detected in and isolated from mosquitoes collected in Haiti, 2016.
Abstract
As part of on-going arboviral surveillance activity in a semi-rural region in Haiti, Chikungunya virus (CHIKV)-positive mosquito pools were identified in 2014 (the peak of the Caribbean Asian-clade epidemic), and again in 2016 by RT-PCR. In 2014, CHIKV was only identified in Aedes aegypti (11 positive pools/124 screened). In contrast, in sampling in 2016, CHIKV was not identified in Ae. aegypti, but, rather, in (a) a female Aedes albopictus pool, and (b) a female Culex quinquefasciatus pool. Genomic sequence analyses indicated that the CHIKV viruses in the 2016 mosquito pools were from the East-Central-South African (ECSA) lineage, rather than the Asian lineage. In phylogenetic studies, these ECSA lineage strains form a new ECSA subgroup (subgroup IIa) together with Brazilian ECSA lineage strains from an isolated human outbreak in 2014, and a mosquito pool in 2016. Additional analyses date the most recent common ancestor of the ECSA IIa subgroup around May 2007, and the 2016 Haitian CHIKV genomes around December 2015. Known CHIKV mutations associated with improved Ae. albopictus vector competence were not identified. Isolation of this newly identified lineage from Ae. albopictus is of concern, as this vector has a broader geographic range than Ae. aegypti, especially in temperate areas of North America, and stresses the importance for continued vector surveillance.
Dollars and antisense for Duchenne muscular dystrophy: Eteplirsen and dystrophin.
Author information
- 1
- From the Department of Neurology (C.D.Z.), University of Pennsylvania, Philadelphia; Division of Pediatric Neurology (P.B.K.), Department of Pediatrics, and Departments of Neurology (P.B.K.), and Molecular Genetics and Microbiology (P.B.K.), University of Florida College of Medicine; and Genetics Institute and Myology Institute (P.B.K.), University of Florida, Gainesville.
- 2
- From the Department of Neurology (C.D.Z.), University of Pennsylvania, Philadelphia; Division of Pediatric Neurology (P.B.K.), Department of Pediatrics, and Departments of Neurology (P.B.K.), and Molecular Genetics and Microbiology (P.B.K.), University of Florida College of Medicine; and Genetics Institute and Myology Institute (P.B.K.), University of Florida, Gainesville. pbkang@ufl.edu.
AAV6 K531 serves a dual function in selective receptor and antibody ADK6 recognition.
Author information
- 1
- Department of Biochemistry & Molecular Biology, Center for Structural Biology, The McKnight Brain Institute, College of Medicine, University of Florida, 1600 SW Archer Road, P.O. Box 100245, Gainesville, FL 32610, USA.
- 2
- Gene Therapy Center and the Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
- 3
- German Cancer Research Center, Heidelberg, Germany.
- 4
- Department of Biochemistry & Molecular Biology, Center for Structural Biology, The McKnight Brain Institute, College of Medicine, University of Florida, 1600 SW Archer Road, P.O. Box 100245, Gainesville, FL 32610, USA. Electronic address: mckenna@ufl.edu.
Abstract
Adeno-associated viruses (AAVs) are being developed as vectors for the treatment of genetic disorders. However, pre-existing antibodies present a significant limitation to achieving optimal efficacy for the AAV gene delivery system. Efforts aimed at engineering vectors with the ability to evade the immune response include identification of residues on the virus capsid important for these interactions and changing them. Here K531 is identified as the determinant of monoclonal antibody ADK6 recognition by AAV6, and not the closely related AAV1. The AAV6-ADK6 complex structure was determined by cryo-electron microscopy and the footprint confirmed by cell-based assays. The ADK6 footprint overlaps previously identified AAV antigenic regions and neutralizes by blocking essential cell surface glycan attachment sites. This study thus expands the available repertoire of AAV-antibody information that can guide the design of host immune escaping AAV vectors able to maintain capsid functionality.
Arthritis gene therapy is becoming a reality.
Author information
- 1
- Rehabilitation Medicine Research Center, Mayo Clinic, Rochester, MN, USA. Evans.Christopher@mayo.edu.
- 2
- Department of Orthopaedics and Rehabilitation, University of Florida College of Medicine, Gainesville, FL, USA.
- 3
- Department of Molecular Medicine, The Scripps Research Institute, Jupiter, FL, USA.
Intramammary 25-hydroxyvitamin D3 treatment modulates innate immune responses to endotoxin-induced mastitis.
Author information
- 1
- Animal Molecular and Cellular Biology Graduate Program, Gainesville 32611.
- 2
- Department of Animal Sciences, University of Florida, Gainesville 32611.
- 3
- Department of Animal Sciences, University of Florida, Gainesville 32611. Electronic address: cdnelson@ufl.edu.
Abstract
Vitamin D signaling in response to pathogen-associated molecules contributes to activation of innate immune responses of bovine monocytes. We hypothesized that lipopolysaccharide (LPS) of bacteria associated with mastitis in dairy cows activates the vitamin D pathway in innate immune cells of the udder and that increasing availability of 25-hydroxyvitamin D3 [25(OH)D3] would augment expression of vitamin D-associated genes. The objective of this experiment was to determine the effects of intramammary LPS and 25(OH)D3 treatments on activation of the vitamin D pathway and innate immune responses of mammary immune cells. Individual mammary quarters of 5 lactating cows were treated with placebo control, 100 μg of 25(OH)D3, 5 μg of LPS, or a combination of 100 μg of 25(OH)D3 and 5 μg of LPS. Somatic cells from milk were evaluated for percentage of neutrophil and macrophage populations and expression of genes associated with vitamin D metabolism and innate immunity. Data from samples collected from 4 to 12 h after challenge were analyzed for main effects of LPS and 25(OH)D3 treatments, treatment interactions, and simple effects of 25(OH)D3 treatment. Data from samples collected at the time of challenge were used as covariates. The percentages of neutrophils in milk at 8 h postchallenge were 58 ± 10, 82 ± 11, 89 ± 10, and 63 ± 10% of total cells in milk from control, 25(OH)D3, LPS, and LPS plus 25(OH)D3 glands, respectively, such that the interaction of LPS and 25(OH)D3 was significant. Expression of the vitamin D 1α-hydroxylase (CYP27B1) and vitamin D receptor genes was upregulated by LPS treatment in total cells, macrophages, and neutrophils in milk. In addition, expression of the vitamin D 24-hydroxylase (CYP24A1) gene in milk somatic cells was upregulated by 25(OH)D3 and LPS treatments. The inducible nitric oxide synthase (iNOS), chemokine (C-C-motif) ligand 5 (CCL5), β-defensin 3 (DEFB3), DEFB7, and DEFB10 genes were upregulated by LPS treatment in total cells and neutrophils from milk. Expression of iNOS in milk somatic cells tended to be affected by the interaction between LPS and 25(OH)D3, such that 25(OH)D3 tended to increase iNOS in the absence of LPS but not in the presence of LPS. Furthermore, expression of CCL5 in macrophages was downregulated by 25(OH)D3. In conclusion, intramammary endotoxin challenge activates the vitamin D pathway in mammary macrophages and neutrophils, and intramammary 25(OH)D3 treatment alters the percentage of neutrophils and expression of immune genes in milk somatic cells.
Neuroscience researches at Belyaev conference-2017.
Author information
- 1
- Novosibirsk State University, Novosibirsk, Russia. orlov@bionet.nsc.ru.
- 2
- The A.O. Kovalevsky Institute of Marine Biological Research of RAS, Sevastopol, Russia. orlov@bionet.nsc.ru.
- 3
- University of Florida, Gainesville, FL, USA.
- 4
- Research Centre of Medical Genetics, Moscow, Russia.
- 5
- Moscow Institute of Physics and Technology (State University), Dolgoprudnyi, Russia.
Functional characterization of the Hyles euphorbiae hawkmoth transcriptome reveals strong expression of phorbol ester detoxification and seasonal cold hardiness genes.
Author information
- 1
- Museum of Zoology, Senckenberg Natural History Collections Dresden, Koenigsbruecker Landstrasse 159, D-01109 Dresden, Germany.
- 2
- 2Florida Museum of Natural History, University of Florida, Gainesville, FL 32611 USA.
- 3
- 3Department of Entomology, China Agricultural University, Bejing, 100193 China.
- 4
- 4China National Gene Bank, Beijing Genomics Institute, Shenzhen, 518083 China.
- 5
- 5GenXPro GmbH, Altenhöferallee 3, D-60438 Frankfurt am Main, Germany.
Abstract
BACKGROUND:
The European spurge hawkmoth, Hyles euphorbiae (Lepidoptera, Sphingidae), has been intensively studied as a model organism for insect chemical ecology, cold hardiness and evolution of species delineation. To understand species isolation mechanisms at a molecular level, this study aims at determining genetic factors underlying two adaptive ecological trait candidates, phorbol ester (TPA) detoxification and seasonal cold acclimation.
METHOD:
A draft transcriptome of H. euphorbiae was generated using Illumina sequencing, providing the first genomic resource for the hawkmoth subfamily Macroglossinae. RNA expression levels in tissues of experimental TPA feeding larvae and cooled pupae was compared to levels in control larvae and pupae using 26 bp RNA sequence tag libraries (DeepSuperSAGE). Differential gene expression was assessed by homology searches of the tags in the transcriptome.
RESULTS:
In total, 389 and 605 differentially expressed transcripts for detoxification and cold hardiness, respectively, could be identified and annotated with proteins. The majority (22 of 28) of differentially expressed detox transcripts of the four ‘drug metabolism’ enzyme groups (cytochrome P450 (CYP), carboxylesterases (CES), glutathione S-transferases (GST) and lipases) are up-regulated. Triacylglycerol lipase was significantly over proportionally annotated among up-regulated detox transcripts. We record several up-regulated lipases, GSTe2, two CESs, CYP9A21, CYP6BD6 and CYP9A17 as candidate genes for further H. euphorbiae TPA detoxification analyses. Differential gene expression of the cold acclimation treatment is marked by metabolic depression with enriched Gene Ontology terms among down-regulated transcripts almost exclusively comprising metabolism, aerobic respiration and dissimilative functions. Down-regulated transcripts include energy expensive respiratory proteins like NADH dehydrogenase, cytochrome oxidase and ATP synthase. Gene expression patterns show shifts in carbohydrate metabolism towards cryoprotectant production. The Glycolysis enzymes, G1Pase, A1e, Gpi and an Akr isoform are up-regulated. Glycerol, an osmolyte which lowers the body liquid supercooling point, appears to be the predominant polyol cryoprotectant in H. euphorbiaediapause pupae. Several protein candidates involved in glucose, glycerol, myo-inositol and potentially sorbitol and trehalose synthesis were identified.
CONCLUSIONS:
A majority of differently expressed transcripts unique for either detoxification or cold hardiness indicates highly specialized functional adaptation which may have evolved from general cell metabolism and stress response.The transcriptome and extracted candidate biomarkers provide a basis for further gene expression studies of physiological processes and adaptive traits in H. euphorbiae.
Recommendations for Laboratory Containment and Management of Gene Drive Systems in Arthropods.
Author information
- 1
- 1 Entomology Branch, Centers for Disease Control and Prevention (CDC), Atlanta, Georgia .
- 2
- 2 Life Sciences, Imperial College London , Ascot, United Kingdom .
- 3
- 3 Department of Parasitology, Institute of Biomedical Sciences, University of Sao Paulo , Sao Paulo, Brazil .
- 4
- 4 National Institute of Science and Technology in Molecular Entomology , National Council of Scientific and Technological Development (INCT-EM/CNPq), Rio de Janeiro, Brazil .
- 5
- 5 CSIRO , Brisbane, Australia .
- 6
- 6 USDA-ARS, Center for Medical, Agricultural, and Veterinary Entomology , Gainesville, Florida.
- 7
- 7 CSIRO , Hobart, Australia .
- 8
- 8 Divisions of Biostatistics and Epidemiology, School of Public Health, University of California , Berkeley, California.
- 9
- 9 Florida Medical Entomology Laboratory, Department of Entomology and Nematology, University of Florida , Vero Beach, Florida.
- 10
- 10 Department of Entomology, Texas A&M University , College Station, Texas.
Abstract
Versatile molecular tools for creating driving transgenes and other invasive genetic factors present regulatory, ethical, and environmental challenges that should be addressed to ensure their safe use. In this article, we discuss driving transgenes and invasive genetic factors that can potentially spread after their introduction into a small proportion of individuals in a population. The potential of invasive genetic factors to increase their number in natural populations presents challenges that require additional safety measures not provided by previous recommendations regarding accidental release of arthropods. In addition to providing physical containment, invasive genetic factors require greater attention to strain management, including their distribution and identity confirmation. In this study, we focus on insects containing such factors with recommendations for investigators who are creating them, institutional biosafety committees charged with ensuring safety, funding agencies providing support, those managing insectaries handling these materials who are responsible for containment, and other persons who will be receiving insects-transgenic or not-from these facilities. We give specific examples of efforts to modify mosquitoes for mosquito-borne disease control, but similar considerations are relevant to other arthropods that are important to human health, the environment, and agriculture.
Regulation of the ATF3 gene by a single promoter in response to amino acid availability and endoplasmic reticulum stress in human primary hepatocytes and hepatoma cells.
Author information
- 1
- Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, Florida 32610.
- 2
- Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, Florida 32610. Electronic address: mkilberg@ufl.edu.
Abstract
Activating transcription factor 3 (ATF3) is a highly regulated protein that is implicated in a wide range of pathological conditions including inflammation and transformation. Transcription from the ATF3 gene is induced by several stress-induced signaling pathways, including amino acid limitation (amino acid response, AAR) and ER stress (unfolded protein response, UPR). Induction of ATF3 transcription by these pathways is mediated by ATF4 and cJUN recruitment to enhancer elements within the ATF3 gene. Although a canonical promoter (promoter A) has been studied by numerous laboratories, a second promoter activity (promoter A1), 43 kb upstream of the first, has been reported to respond to stress-induced signaling and to be critical for ATF3 expression in certain transformed cells. The results of the present study show that in normal human hepatocytes and HepG2 human hepatoma cells both basal as well as AAR- and UPR-induced transcription occurs almost exclusively from promoter A. This selectivity between the two promoters correlated with increased binding of ATF4, recruitment of RNA polymerase II, and the expected histone modifications in the promoter A region of the gene. Time course studies of ATF3 transcription activity revealed that the temporal kinetics for ATF3 induction differ between the AAR and UPR, with the former being more transient than the latter. Collectively, the results document that ATF3 expression in normal and transformed human liver originates from the canonical promoter A that responds to multiple stress signals.
Dicyanomethylene Substituted Benzothiazole Squaraines: The Efficiency of Photodynamic Therapy In Vitro and In Vivo.
Author information
- 1
- Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China; Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Guangxi Medical University, No. 22, Shuangyong Road, Nanning 530021, Guangxi, PR China.
- 2
- Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Biology and College of Chemistry and Chemical Engineering, Hunan University Changsha, 410082, China.
- 3
- Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China; University of the Chinese Academy of Sciences, Beijing 100049, China.
- 4
- Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
- 5
- Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Biology and College of Chemistry and Chemical Engineering, Hunan University Changsha, 410082, China; Department of Chemistry, Center for Research at the Bio/Nano Interface, Health Cancer Center, UF Genetics Institute, McKnight Brain Institute, University of Florida Gainesville, FL 32611-7200, USA. Electronic address: tan@chem.ufl.edu.
- 6
- Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China; University of the Chinese Academy of Sciences, Beijing 100049, China. Electronic address: sgdh@iccas.ac.cn.
Abstract
The lack of ideal photosensitizers limits the clinicalapplication of photodynamic therapy (PDT). Here we report the PDT efficiency of dicyanomethylene substituted benzothiazole squaraine derivatives. This class of squaraine derivatives possess strong absorption and long excitation and emission wavelengths (ex/em, 685/720nm). They show negligible dark toxicity, but can generate singlet oxygen under irradiation resulting in the apoptosis and necrosis of cells (phototoxicity). Changing the side chains of these compounds greatly influences their albumin-binding rate, cellular uptake and their phototoxicity. One of the squaraine derivatives with two methyl butyrate side chains shows high PDT efficiency in a mouse subcutaneous xenograft model under the irradiation of a 690nm laser. These results show the great potential of dicyanomethylene substituted benzothiazole squaraines to be the leading compound of near-infrared photosensitizers in PDT.