ABCG Family Research Articles

Transcriptome mining and expression analysis of ABC transporter genes in a monophagous herbivore, Leucinodes orbonalis (Crambidae: Lepidoptera)

Insecticide resistance is a global concern and requires immediate attention to manage dreadful insect pests. One of the resistance mechanisms adopted by insects is through ATP-binding cassette (ABC) transporter proteins. These proteins rapidly transport and eliminate the insecticidal molecules across the lipid membranes (Phase III detoxification mechanism). In the present study, we investigated the potential role of ABC transporter genes in imparting insecticide resistance in field-collected insecticide resistant larvae of eggplant shoot and fruit borer (Leucinodes orbonalis; Crambidae: Lepidoptera). Dose-mortality bioassays against five insecticidal molecules revealed moderate to high levels of insecticide resistance (32.2. to 134.1-fold). Thirty-one genes encoding ABC transporter proteins were mined from the transcriptome resources of L. orbonalis. They were classified under eight sub-families (ABCA to ABCH). Phylogenetic analysis indicated ABCG is the most divergent, composed of nine genes as compared to many other insects. Transcriptome analysis of the insecticide resistant and susceptible strains of L. orbonalis revealed differential expression of 13 ABC transporter genes. The altered expression of these genes was further validated using qRT-PCR. The knockdown studies indicated the involvement of ABCD1 and ABCG2 genes in chlorantraniliprole resistance in the insecticide-resistant strain of L. orbonalis. This study unveils the additional mechanisms employed by L. orbonalis in resisting insecticide toxicity through accelerated excretion mode. These ABCD and ABCG family genes could be candidate targets in developing genome-assisted pest management strategies in the future.

Comparative Transcriptome and Metabolome Analyses Revealed Quality Difference between Beauty Tea Processed through Indoor Withering and Outdoor Solar Withering.

Withering is the first processing procedure of beauty tea, and there are few reports on the impact of withering methods on the quality of beauty tea and its regulatory mechanisms. Through comparison of fresh tea leaves (FT) with the leaves at Indoor natural Withering 18h (IWT-18) and Outdoor Solar Withering 6h (OWT-6), which were collected at the end of the two withering processes, 17282 and 13984 differentially expressed genes (DEGs) were screened and 267 and 154 differential metabolites (DMs) were identified, respectively. Coexpression network revealed that a large number of DEGs and DMs were enriched in phenylpropanoid, flavonoid, and adenosine Triphosphate binding (ABC) transporter pathways, and the number of DMs and DEGs in IWT-18 vs FT exceeded that in OWT-6 vs FT. Both withering methods promoted a significant increase in content of phenylalanine and upregulation of β-glucoside (BGLU) expression in the phenylpropanoid metabolism pathway. The five proanthocyanidins related to theaflavins in the flavonoid synthesis pathway was more significant accumulation in FT vs IWT-18 than in FT vs OWT-6. Meanwhile, both withering methods can affect the ABC transporter pathway to promote the accumulation of amino acids and their derivatives, but different withering methods affect different ABC transporter families. Outdoor withering with more severe abiotic stress has a greater impact on the ABCG family, while indoor withering has a more significant effect on the ABCC family. Sensory evaluation results showed that the dry tea of IWT-18 was slightly better than that of OWT-6 because of longer withering time and more thorough substance transformation. In conclusion, the formation of honey flavor in beauty tea may be closely related to the DEGs and DMs in these three pathways. Our research provides theoretical data support for further revealing the mechanism of quality formation during the withering process of beauty tea. This article is protected by copyright. All rights reserved.

ATP-Binding Cassette (ABC) Transporters in Fusarium Specific Mycoparasite Sphaerodes mycoparasitica during Biotrophic Mycoparasitism

Recent transcriptomic profiling has revealed importance membrane transporters such as ATP-binding cassette (ABC) transporters in fungal necrotrophic mycoparasites. In this study, RNA-Seq allowed rapid detection of ABC transcripts involved in biotrophic mycoparasitism of Sphaerodes mycoparasitica against the phytopathogenic and mycotoxigenic Fusarium graminearum host, the causal agent of Fusarium head blight (FHB). Transcriptomic analyses of highly expressed S. mycoparasitica genes, and their phylogenetic relationships with other eukaryotic fungi, portrayed the ABC transporters’ evolutionary paths towards biotrophic mycoparasitism. Prior to the in silico phylogenetic analyses, transmission electron microscopy (TEM) was used to confirm the formation of appressorium/haustorium infection structures in S. mycoparasitica during early (1.5 d and 3.5 d) stages of mycoparasitism. Transcripts encoding biotrophy-associated secreted proteins did uncover the enrolment of ABC transporter genes in this specific biocontrol mode of action, while tandem ABC and BUB2 (non-ABC) transcripts seemed to be proper for appressorium development. The next-generation HiSeq transcriptomic profiling of the mycoparasitic hypha samples, revealed 81 transcripts annotated to ABC transporters consisting of a variety of ABC-B (14%), ABC-C (22%), and ABC-G (23%), and to ABC-A, ABC-F, aliphatic sulfonates importer (TC 3.A.1.17.2), BtuF, ribose importer (TC 3.A.1.2.1), and unknown families. The most abundant transcripts belonged to the multidrug resistance exporter (TC 3.A.1.201) subfamily of the ABC-B family, the conjugate transporter (TC 3.A.1.208) subfamily of the ABC-C family, and the pleiotropic drug resistance (PDR) (TC 3.A.1.205) subfamily of the ABC-G family. These findings highlight the significance of ABC transporter genes that control cellular detoxification against toxic substances (e.g., chemical pesticides and mycotoxins) in sustaining a virulence of S. mycoparasitica for effective biotrophic mycoparasitism on the F. graminearum host. The findings of this study provide clues to better understand the biotrophic mycoparasitism of S. mycoparasitica interacting with the Fusarium host, which implies that the ABC transporter group of key proteins is involved in the mycoparasite’s virulence and multidrug resistance to toxic substances including cellular detoxification.

The AbcCl1 transporter of Colletotrichum lindemuthianum acts as a virulence factor involved in fungal detoxification during common bean (Phaseolus vulgaris) infection.

Anthracnose, caused by Colletotrichum lindemuthianum, is a disease affecting the common bean plant, Phaseolus vulgaris. To establish infection, the phytopathogen must survive the toxic compounds (phytoanticipins and phytoalexins) that are produced by the plant as a defense mechanism. To study the detoxification and efflux mechanisms in C. lindemuthianum, the abcCl1 gene, which encodes an ABC transporter, was analyzed. The abcCl1 gene (4558 pb) was predicted to encode a 1450-amino acid protein. Structural analysis of 11 genome sequences from Colletotrichum spp. showed that the number of ABC transporters varied from 34 to 64. AbcCl1 was classified in the ABC-G family of transporters, and it appears to be orthologs to ABC1 from Magnaporthe grisea and FcABC1 from Fusarium culmorum, which are involved in pleiotropic drug resistance. A abcT3 (ΔabcCl1) strain showed reduction on aggressivity when inoculated on bean leaves that presented diminishing anthracnose symptoms, which suggests the important role of AbcCl1 as a virulence factor and in fungal resistance to host compounds. The expression of abcCl1 increased in response to different toxic compounds, such as eugenol, hygromycin, and pisatin phytoalexin. Together, these results suggest that AbcCl1 is involved in fungal resistance to the toxic compounds produced by plants or antagonistic microorganisms.

Functional analysis of cyantraniliprole tolerance ability mediated by ATP-binding cassette transporters in Aphis gossypii glover

Cyantraniliprole, a second-generation anthranilic diamide insecticide, is widely used to control chewing and sucking pests. ATP-binding cassette transporters (ABCs) are a ubiquitous family of membrane proteins that play important roles in insect detoxification mechanisms. However, the potential effects of ABCs on cyantraniliprole-resistance remain unclear. In the present study, synergism bioassays revealed that verapamil, an ABC inhibitor, increased the toxicity of cyantraniliprole by 2.00- and 12.25-fold in the susceptible and cyantraniliprole-resistant strains of Aphis gossypii. Based on transcriptome data, the expression levels of ABCB4, ABCB5, ABCD1, ABCG4, ABCG7, ABCG13, ABCG16, ABCG17, ABCG26 and MRP12 were upregulated 1.56-, 1.32-, 1.51-, 2.03-, 1.65-, 1.50-, 4.18-, 6.07-, 4.68- and 4.69-fold, respectively, in the cyantraniliprole-resistant strain (CyR) compared to the susceptible strain (SS), as determined using RT-qPCR. Drosophila melanogaster ectopically overexpressing ABCB5, ABCG4, ABCG7, ABCG16, ABCG17, ABCG26 and MRP12 exhibited significantly increased tolerance to cyantraniliprole by 11.71-, 2.39-, 4.85-, 2.06-, 3.75-, 4.20- and 3.50-fold, respectively, with ABCB5 and ABCG family members being the most effective. Furthermore, the suppression of ABCB5, ABCG4, ABCG7, ABCG16, ABCG17, ABCG26 and MRP12 significantly increased the sensitivity of the CyR strain to cyantraniliprole. These results indicate that ABCs may play crucial roles in cyantraniliprole resistance and may provide information for shaping resistance management strategies.

An ABCG Transporter Functions in Rab Localization and Lysosome-Related Organelle Biogenesis in Caenorhabditis elegans.

ABC transporters couple ATP hydrolysis to the transport of substrates across cellular membranes. This protein superfamily has diverse activities resulting from differences in their cargo and subcellular localization. Our work investigates the role of the ABCG family member WHT-2 in the biogenesis of gut granules, a Caenorhabditis elegans lysosome-related organelle. In addition to being required for the accumulation of birefringent material within gut granules, WHT-2 is necessary for the localization of gut granule proteins when trafficking pathways to this organelle are partially disrupted. The role of WHT-2 in gut granule protein targeting is likely linked to its function in Rab GTPase localization. We show that WHT-2 promotes the gut granule association of the Rab32 family member GLO-1 and the endolysosomal RAB-7, identifying a novel function for an ABC transporter. WHT-2 localizes to gut granules where it could play a direct role in controlling Rab localization. Loss of CCZ-1 and GLO-3, which likely function as a guanine nucleotide exchange factor (GEF) for GLO-1, lead to similar disruption of GLO-1 localization. We show that CCZ-1, like GLO-3, is localized to gut granules. WHT-2 does not direct the gut granule association of the GLO-1 GEF and our results point to WHT-2 functioning differently than GLO-3 and CCZ-1 Point mutations in WHT-2 that inhibit its transport activity, but not its subcellular localization, lead to the loss of GLO-1 from gut granules, while other WHT-2 activities are not completely disrupted, suggesting that WHT-2 functions in organelle biogenesis through transport-dependent and transport-independent activities.

Cloning and expression of selected ABC transporters from the Arabidopsis thaliana ABCG family in Pichia pastoris.

Phytohormones play a major role in plant growth and development. They are in most cases not synthesized in their target location and hence need to be transported to the site of action, by for instance ATP-binding cassette transporters. Within the ATP-binding cassette transporter family, Pleiotropic Drug Resistance transporters are known to be involved in phytohormone transport. Interestingly, PDRs are only present in plants and fungi. In contrast to fungi, there are few biochemical studies of plant PDRs and one major reason is that suitable overexpression systems have not been identified. In this study, we evaluate the expression system Pichia pastoris for heterologous overexpression of PDR genes of the model plant Arabidopsis thaliana. We successfully cloned and expressed the potential phytohormone transporters PDR2 and PDR8 in P. pastoris. Sucrose gradient centrifugation confirmed that the overexpressed proteins were correctly targeted to the plasma membrane of P. pastoris and initial functional studies demonstrated ATPase activity for WBC1. However, difficulties in cloning and heterologous overexpression might be particular obstacles of the PDR family, since cloning and overexpression of White Brown Complex 1, a half-size transporter of the same ABCG subfamily with comparable domain organization, was more easily achieved. We present strategies and highlight critical factors to successfully clone plant PDR genes and heterologously expressed in P. pastoris.

Information theoretic measures and mutagenesis identify a novel linchpin residue involved in substrate selection within the nucleotide-binding domain of an ABCG family exporter Cdr1p

ABC transporters are membrane-bound pumps composed of two major domains: the transmembrane domain(s) (TMDs) and the nucleotide-binding domain(s) (NBDs). Sequence analyses of the NBDs of key ABC exporters revealed a residue position within the H-loop to be differentially conserved in the ABCG family, wherein there lies glutamine instead of positively charged arginine/lysine as in non-ABCG members. Consequently, contrasting NBD sequences of fungal Pleiotropic Drug Resistance transporters (PDR/ABCG) with that of Cholesterol/Phospholipid and Retinal (CPR/ABCA) Flippase family revealed a high Cumulative Relative Entropy (CRE) score of this residue position implying its family-specific functional significance. Further, substitution of the glutamine by arginine in both the NBDs of a representative PDR/ABCG member, (Candida drug resistance 1 protein) Cdr1p led to selective susceptibility of the Saccharomyces cerevisiae strains overexpressing the corresponding mutant proteins (Q362R and Q1060R) towards antifungal substrates without any impact on the ATPase activity. Consistent with the findings from previous studies on H-loop motif of fungal PDR transporters, the current report points towards a role of the glutamine residue within both canonical and divergent H-loop of Cdr1p in conferring substrate selection in a precisely identical manner.

OsABCG9 Is an Important ABC Transporter of Cuticular Wax Deposition in Rice.

The importance of the cuticular layer in regulating a plant’s water status and providing protection from environmental challenges has been recognized for a long time. The cuticular layer in plants restricts non-stomatal water loss and protects plants against damage from pathogen infection and UV radiation. Much genetic and biochemical research has been done about cutin and wax transportation in Arabidopsis thaliana, but little is known about it in rice. Here, we report that a rice ATP-binding cassette (ABC) transporter, OsABCG9, is essential for normal development during vegetative growth and could play a critical role in the transportation of epicuticular wax in rice. Rice phenotypes with mutated OsABCG9 exhibited growth retardation and sensitivity to low humidity. The total amount of cuticular wax on the leaves of the osabcg9-1 mutant diminished by 53% compared with the wild type, and wax crystals disappeared completely in osabcg9-2 mutant leaves. However, OsABCG9 does not seem to be involved in cutin transportation, even though its ortholog in Arabidopsis, AtABCG11, transports both wax and cutin. Furthermore, the osabcg9-1 mutant had increased leaf chlorophyll leaching and more severe drought susceptibility. This study provides new insights about differences between rice and A. thaliana in wax and cutin transportation associated with the ABCG family during evolution.

Structure-function relationships in ABCG2: insights from molecular dynamics simulations and molecular docking studies

Efflux pumps of the ATP-binding cassette transporters superfamily (ABC transporters) are frequently involved in the multidrug-resistance (MDR) phenomenon in cancer cells. Herein, we describe a new atomistic model for the MDR-related ABCG2 efflux pump, also named breast cancer resistance protein (BCRP), based on the recently published crystallographic structure of the ABCG5/G8 heterodimer sterol transporter, a member of the ABCG family involved in cholesterol homeostasis. By means of molecular dynamics simulations and molecular docking, a far-reaching characterization of the ABCG2 homodimer was obtained. The role of important residues and motifs in the structural stability of the transporter was comprehensively studied and was found to be in good agreement with the available experimental data published in literature. Moreover, structural motifs potentially involved in signal transmission were identified, along with two symmetrical drug-binding sites that are herein described for the first time, in a rational attempt to better understand how drug binding and recognition occurs in ABCG2 homodimeric transporters.

Identification of ABC transporters acting in vitamin B12 metabolism in Caenorhabditis elegans.

Vitamin B12 (cobalamin, Cbl) is a micronutrient essential to human health. Cbl is not utilized as is but must go through complex subcellular and metabolic processing to generate two cofactor forms: methyl-Cbl for methionine synthase, a cytosolic enzyme; and adenosyl-Cbl for methylmalonyl-CoA mutase, a mitochondrial enzyme. Some 10-12 human genes have been identified responsible for the intracellular conversion of Cbl to cofactor forms, including genes that code for ATP-binding cassette (ABC) transporters acting at the lysosomal and plasma membranes. However, the gene for mitochondrial uptake is not known. We hypothesized that ABC transporters should be candidates for other uptake and efflux functions, including mitochondrial transport, and set out to screen ABC transporter mutants for blocks in Cbl utilization using the nematode roundworm Caenorhabditis elegans. Thirty-seven mutant ABC transporters were screened for the excretion of methylmalonic acid (MMA), which should result from loss of Cbl transport into the mitochondria. One mutant, wht-6, showed elevated MMA excretion and reduced [14C]-propionate incorporation, pointing to a functional block in methylmalonyl-CoA mutase. In contrast, the wht-6 mutant appeared to have a normal cytosolic pathway based on analysis of cystathionine excretion, suggesting that cytosolic methionine synthase was functioning properly. Further, the MMA excretion in wht-6 could be partially reversed by including vitamin B12 in the assay medium. The human ortholog of wht-6 is a member of the G family of ABC transporters. We propose wht-6 as a candidate for the transport of Cbl into mitochondria and suggest that a member of the corresponding ABCG family of ABC transporters has this role in humans. Our ABC transporter screen also revealed that mrp-1 and mrp-2 mutants excreted lower MMA than wild type, suggesting they were concentrating intracellular Cbl, consistent with the cellular efflux defect proposed for the mammalian MRP1 ABC transporter.

A major QTL controlling apple skin russeting maps on the linkage group 12 of 'Renetta Grigia di Torriana'.

BackgroundRusseting is a disorder developed by apple fruits that consists of cuticle cracking followed by the replacement of the epidermis by a corky layer that protects the fruit surface from water loss and pathogens. Although influenced by many environmental conditions and orchard management practices, russeting is under genetic control. The difficulty in classifying offspring and consequent variable segregation ratios have led several authors to conclude that more than one genetic determinant could be involved, although some evidence favours a major gene (Ru).ResultsIn this study we report the mapping of a major genetic russeting determinant on linkage group 12 of apple as inferred from the phenotypic observation in a segregating progeny derived from ‘Renetta Grigia di Torriana’, the construction of a 20 K Illumina SNP chip based genetic map, and QTL analysis. Recombination analysis in two mapping populations restricted the region of interest to approximately 400 Kb. Of the 58 genes predicted from the Golden Delicious sequence, a putative ABCG family transporter has been identified. Within a small set of russeted cultivars tested with markers of the region, only six showed the same haplotype of ‘Renetta Grigia di Torriana’.ConclusionsA major determinant (Ru_RGT) for russeting development putatively involved in cuticle organization is proposed as a candidate for controlling the trait. SNP and SSR markers tightly co-segregating with the Ru_RGT locus may assist the breeder selection. The observed segregations and the analysis of the ‘Renetta Grigia di Torriana’ haplotypic region in a panel of russeted and non-russeted cultivars may suggest the presence of other determinants for russeting in apple.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0507-4) contains supplementary material, which is available to authorized users.

A half-type AvABCG1 transporter derived from Andropogon virginicus L. confers aluminum tolerance

Aluminum (Al) induced genes were isolated from a wild plant, Andropogon virginicus L. by a method involving arbitrarily primed PCR-based RNA finger printing. One of the clones, AP18-2-1, was induced in both roots and shoots by Al and diamide treatments, and slightly by Cu and Zn treatments. Full-length cDNA (2287bp) was obtained by RACE (rapid amplification of cDNA ends) PCR and DNA sequencing indicated that it encoded a half-type ABC transporter that was highly homologous to other ABCG family genes. The Al-induced transporter gene derived from A. virginicus L. was therefore designated AvABCG1. The AvABCG1 gene conferred tolerance to Al, Cu and diamide in Arabidopsis thaliana transformants. A fusion protein (AvABCG1::GFP) was localized in the cell membrane region. Immunohistological analysis of the native protein in A. virginicus L. indicated that it localized to both exodermis and endodermis in roots, and to the vascular bundle sheath and its extensions in both upper and lower epidermal cells in leaves. Fluorescent measurement showed that absorbed Al was detected in almost the same areas as the AvABCG1 protein in both root and leaf. We conclude that the AvABCG1 transporter may confer Al tolerance by accumulating absorbed Al ions in specific regions of the plant.

Effect of nine diets on xenobiotic transporters in livers of mice

1. Lifestyle diseases are often caused by inappropriate nutrition habits and attempted to be treated by polypharmacotherapy. Therefore, it is important to determine whether differences in diet affect the disposition of drugs. Xenobiotic transporters in the liver are essential in drug disposition.2. In the current study, mice were fed one of nine diets for 3 weeks. The mRNAs of 23 known xenobiotic transporters in livers of mice were quantified by microarray analysis, and validated by branched DNA assay. The mRNAs of 15 transporters were altered by at least one diet. Diet-restriction (10) and the atherogenic diet (10) altered the expression of the most number of transporters, followed by western diet (8), high-fat diet (4), lab chow (2), high-fructose diet (2) and EFA-deficient diet (2), whereas the low n−3 FA diet had no effect on these transporters. Seven of the 11 xenobiotic transporters in the Slc family, three of four in the Abcb family, two of four in the Abcc family and all three in the Abcg family were changed significantly.3. This first comprehensive study indicates that xenobiotic transporters are altered by diet, and suggests there are likely diet–drug interactions due to changes in the expression of drug transporters.

Identification and differential induction of ABCG transporter genes in wheat cultivars challenged by a deoxynivalenol-producing Fusarium graminearum strain

Fusarium head blight (FHB), predominantly caused by Fusarium graminearum, is a devastating disease that poses a serious threat to wheat (Triticum aestivum L.) production worldwide. A suppression subtractive hybridization cDNA library was constructed from F. graminearum infected spikes of a resistant Belgian winter wheat, Centenaire, exhibiting Type II resistance to FHB in order to identify differentially expressed members of full-size ABCG family. Members of the ABCG family are pleiotropic drug transporters allowing the movement of structurally unrelated metabolites, including pathogens-derived virulent compounds, across biological membranes and could be potentially involved in resistance to plant pathogens. In this study, five new full-size ABCG transporter expressed sequence tags TaABCG2, TaABCG3, TaABCG4, TaABCG5 and TaABCG6 have been identified. Time-course gene expression profiling between the FHB resistant Centenaire and the susceptible Robigus genotype showed that the newly isolated transcripts were differentially expressed up to 72 h-post inoculation. The respective genes encoding these transcripts were mapped to corresponding wheat chromosomes or chromosomal arms known to harbor quantitative trait loci for FHB resistance. Interestingly, these ABCG transcripts were also induced by deoxynivalenol (DON) treatment of germinating wheat seeds and the toxin treatment inhibited root and hypocotyl growth. However, the hypocotyl of the FHB resistant cultivar Centenaire was less affected than that of the susceptible cultivar Robigus, reflecting more likely the genotype-dependent differential expression pattern of the identified ABCG genes. This work emphasizes the potential involvement of ABCG transporters in wheat resistance to FHB, at least in part through the detoxification of the pathogen-produced DON.

Expression pattern of human ATP-binding cassette transporters in skin.

ATP-binding cassette (ABC) transporters transport a variety of substrates across cellular membranes coupled with hydrolysis of ATP. Currently 49 ABC transporters consisting of seven subfamilies, ABCA, ABCB, ABCC, ABCD, ABCE, ABCF, and ABCG, have been identified in humans and they are extensively expressed in various tissues. Skin can develop a number of drug-induced toxicities' such as Stevens–Johnson syndrome and psoriasis. Concentration of drugs in the skin cells is associated with the development of adverse drug reactions. ABC transporters play important roles in absorption and disposition of drugs in the cells; however, the expression pattern of human ABC transporters in the skin has not been determined. In this study, the expression patterns of 48 human ABC transporters were determined in the human skin as well as in the liver and small intestine. Most of the ABCA, ABCB, ABCC, ABCD, ABCE, and ABCF family members were highly or moderately expressed in the skin, while ABCG family members were slightly expressed in the skin. Significant interindividual variability was also observed in the expression levels of those ATP transporters in the skin, except for ABCA5 and ABCF1, which were found to be expressed in all of the human skin samples tested in this study. In conclusion, this is the first study to identify the expression pattern of the whole human ABC family of transporters in the skin. The interindividual variability in the expression levels of ABC transporters in the human skin might be associated with drug-induced skin diseases.

ABCG8 Gene Responses to 8 Weeks Treadmill Running With or Without Pistachia atlantica (Baneh) Extraction in Female Rats

BackgroundIt is well established that the excess cellular cholesterol concentration, as well as high density lipoprotein (HDL) and total cholesterol levels are strongly correlated with the incidence of coronary artery disease (CAD). Reverse cholesterol transport (RCT) is a term used to describe the efflux of excess cellular cholesterol. ABCG8 is a member of ABCG family that play a critical role in this process.ObjectivesThe current study was conducted to investigate the effect of endurance exercise with or without Pistachia atlantica (Baneh) extraction on small intestine and kidney ABCG8 gene, also plasma high density lipoprotein (HDL-c), triglyceride (TG), total cholesterol (TC), glucose, and estrogen levels in female rats.Materials and MethodsIn this study twenty Wistar female rats (six to eight weeks old, 125-135 g weight) were used. Animals were randomly assigned into training (n = 10) and control (n = 10) groups and further divided into saline-control (SC), saline-training (ST), Baneh-control (BC), and Baneh-training (BT) groups. Training groups was given exercise on a motor-driven treadmill at 25 m/min (0% grade) for 60 min/day, 5 days/week for eight weeks. Animals were fed orally with Baneh extraction and saline for four week. After the last training session, rats were sacrificed, small intestine and kidney were excised, and ABCG8 expression was detected by Real-time PCR method. Plasma also was collected for plasma variable measurements. Statistical analysis was performed using a one way analysis of variance, and significance was accepted at P < 0.05. Correlation was calculated using the Pearson Product Moment correlation.ResultsExercise increased (P < 0.01) and Baneh reduced intestinal ABCG8 mRNA (P < 0.05). In kidney tissue, there wasn’t significant change between the groups (P < 0.40). Plasma HDL-C level was increased by exercise (P < 0.05) and decreased by Baneh (P < 0.02) that was correlated by intestine ABCG8 (r = 0.81, P < 0.001). Plasma TG and TC were unchanged, but glucose and estradiol were increased and decreased in Baneh groups (P < 0.02), respectively.ConclusionsOur study shows that exercise increases intestinal ABCG8 mRNA, and Baneh can increase plasma glucose concentration and reduce ABCG8 expression, HDL-C, and estrogen levels probably due to high fatty acid components.

The ABCG family of membrane‐associated transporters: you don't have to be big to be mighty

Along with many other mammalian ATP-binding cassette (ABC) transporters, members of the ABCG group are involved in the regulated transport of hydrophobic compounds across cellular membranes. In humans, five ABCG family members have been identified, encoding proteins ranging from 638 to 678 amino acids in length. All five have been the subject of intensive investigation to better understand their physiological roles, expression patterns, interactions with substrates and inhibitors, and regulation at both the transcript and protein level. The principal substrates for at least four of the ABCG proteins are endogenous and dietary lipids, with ABCG1 implicated in particular in the export of cholesterol, and ABCG5 and G8 forming a functional heterodimer responsible for plant sterol elimination from the body. ABCG2 has a much broader substrate specificity and its ability to transport numerous diverse pharmaceuticals has implications for the absorption, distribution, metabolism, excretion and toxicity (ADMETOx) profile of these compounds. ABCG2 is one of at least three so-called multidrug resistant ABC transporters expressed in humans, and its activity is associated with decreased efficacy of anti-cancer agents in several carcinomas. In addition to its role in cancer, ABCG2 also plays a role in the normal physiological transport of urate and haem, the implications of which are described. We summarize here data on all five human ABCG transporters and provide a current perspective on their roles in human health and disease.

Transporters are an under‐developed therapeutic target. Discuss

Transporters are an under‐developed therapeutic target. Discuss

The ABC of ABCs: multidrug resistance and genetic diseases.

ATP-binding cassette (ABC) proteins comprise one the largest protein families, because thousands of distinct genes are known to operate in all living cells and organisms from bacteria to man. Most ABC proteins act as transporters, required for a remarkable variety of membrane translocation processes through both efflux and uptake. ABC substrates include ions, heavy metals, carbohydrates, amino acids, oligopeptides, steroids, glucocorticoids, phospholipids, cholesterol, mycotoxins, antibiotics, drugs, pigments, peptide pheromones and even large proteins. Most strikingly, ABC proteins also function as receptors, ion channels, regulators of channels, nutrient sensors and even membrane-bound proteases. However, the mechanism(s) by which evolutionary and structurally conserved ABC proteins can fulfill such functional diversity remains an intriguing and as yet unsolved mystery. The P-glycoprotein (P-gp), multidrug resistancerelated protein (MRP) and breast cancer resistance protein (BCRP) subfamilies have been implicated in multidrug resistance (MDR) phenomena in cancer. Similarly, many orthologs in parasites, fungi and bacteria mediate anti-infective resistance. The clinical significance and contribution of P-gp to antitumor resistance, as well as approaches to overcome MDR in tumors by specific P-gp inhibitors, is more than ever a matter of intense dispute. What stands undisputed is that defective human ABC proteins cause important genetic disorders such as cystic fibrosis, macular dystrophy, hepatic cholestasis, the familial hyperinsulinemic hypoglycemia of infancy, lung dysfunction, connective tissue degeneration, atherosclerosis and even gout. This medical relevance of ABC proteins prompted the first FEBS Advanced Lecture Course in 1997 in Austria. Ever since, biennial FEBS courses, and then the first FEBS Special Meeting on ABC Proteins in 2006, have followed. ABC2010, organized by Kazumitsu Ueda, Susan PC Cole, Susan Michaelis and Karl Kuchler, was the latest in this successful series. In fact, the Austrian ABC meeting has become the most recognized gathering point for the ABC community, and many key collaborations in the field have originated in Innsbruck. Despite intense research efforts in basic and clinical research, as well as in drug discovery, a molecular understanding of the function mechanism of a single human ABC transporter appears elusive. The technical difficulties of studying intrinsic membrane proteins offer one explanation, the painfully slow process in getting high-resolution crystal structures is another. A few elegant, and in some instances debated, structures of ABC transporters have emerged, but the field needs many more (dynamic) structures to unravel common as well as ABC-protein-specific principles of function. FEBS and many other sponsors recognize the importance of the field and provided generous support for ABC2010; they will do so also for ABC2012, the next ABC meeting to take place in Innsbruck (http:// www.febs-abc2012.org). This minireview series, which is based on some exciting findings presented at ABC2010, discusses state-ofthe-art knowledge of hallmark members of the ABCA, ABCC and ABCG families. Two reviews address ABCA1, which connects lipoprotein metabolism with inborn errors of cholesterol transport, and ABCA4, which is tightly linked to hereditary vision loss owing to its role in membrane lipid homeostasis and retinal recycling. Like P-gp, certain ABCC and ABCG members share a remarkable functional dichotomy. On the one hand, they are potentially detrimental, because their ectopic or misregulated expression causes clinical MDR in many malignant tissues. On the other hand, they are essential for physiological drug detoxification processes, especially in the liver, kidney, placenta, across the blood–brain barrier or in the intestinal lumen. Thus, another minireview addresses the ABCG family, in which the physiological substrate for ABCG2, urate, has been recently identified, disclosing its essential role in gout. The fourth minireview discusses ABCC members like the MRP transporters, which are implicated in clinical drug resistance, as well as drug absorption, deposition and hepatic detoxification.