scholarly journals Structure of MlaFB uncovers novel mechanisms of ABC transporter regulation

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Ljuvica R Kolich ◽  
Ya-Ting Chang ◽  
Nicolas Coudray ◽  
Sabrina I Giacometti ◽  
Mark R MacRae ◽  
...  
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Membrane Integrity ◽  
Regulatory Mechanisms ◽  
Nucleotide Binding Domain ◽  
E Coli ◽  
Small Proteins ◽  
Transporter Family ◽  
Bacterial Lipid ◽  
Lipid Transporter

ABC transporters facilitate the movement of diverse molecules across cellular membranes, but how their activity is regulated post-translationally is not well understood. Here we report the crystal structure of MlaFB from E. coli, the cytoplasmic portion of the larger MlaFEDB ABC transporter complex, which drives phospholipid trafficking across the bacterial envelope to maintain outer membrane integrity. MlaB, a STAS domain protein, binds the ABC nucleotide binding domain, MlaF, and is required for its stability. Our structure also implicates a unique C-terminal tail of MlaF in self-dimerization. Both the C-terminal tail of MlaF and the interaction with MlaB are required for the proper assembly of the MlaFEDB complex and its function in cells. This work leads to a new model for how an important bacterial lipid transporter may be regulated by small proteins, and raises the possibility that similar regulatory mechanisms may exist more broadly across the ABC transporter family.

scholarly journals Structure of MlaFB uncovers novel mechanisms of ABC transporter regulation

2020 ◽  
Author(s):  
Ljuvica Kolich ◽  
Ya-Ting Chang ◽  
Nicolas Coudray ◽  
Sabrina I. Giacometti ◽  
Mark R. MacRae ◽  
...  
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Atp Hydrolysis ◽  
Nucleotide Binding ◽  
Binding Domain ◽  
Regulatory Subunit ◽  
Nucleotide Binding Domain ◽  
Transporter Family ◽  
Bacterial Lipid ◽  
Phospholipid Transport

ABC transporters facilitate the movement of a diverse array of molecules across cellular membranes, using power from ATP hydrolysis. While the overall mechanism of the transport cycle has been characterized in detail for several important members of this transporter family, it is less well understood how the activity of ABC transporters is regulated in the cell post-translationally. Here we report the X-ray crystal structure of MlaFB from E. coli, an ABC nucleotide binding domain (MlaF) in complex with its putative regulatory subunit (MlaB). MlaFB constitutes the cytoplasmic portion of the larger MlaFEDB ABC transporter complex, which drives phospholipid transport across the bacterial envelope and is important for maintaining the integrity of the outer membrane barrier. Our data show that the regulatory subunit MlaB, a STAS domain protein, binds to the nucleotide binding domain and is required for its stability. Our structure also implicates a unique C-terminal tail of the ABC subunit, MlaF, in self-dimerization. Both the C-terminal tail of MlaF and the interaction with MlaB are required for the proper assembly of the MlaFEDB complex and its function in cells. This work leads to a new model for how the activity of an important bacterial lipid transporter may be regulated by small binding proteins, and raises the possibility that similar regulatory mechanisms may exist more broadly across the ABC transporter family, from bacteria to humans.

scholarly journals Structural and functional insights into the Diabrotica virgifera virgifera ATP-binding cassette transporter gene family

BMC Genomics ◽  
2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Folukemi Adedipe ◽  
Nathaniel Grubbs ◽  
Brad Coates ◽  
Brian Wiegmman ◽  
Marcé Lorenzen
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Sequence Data ◽  
Transcriptome Assembly ◽  
Diabrotica Virgifera Virgifera ◽  
Atp Binding ◽  
Pest Species ◽  
Diabrotica Virgifera ◽  
Transporter Family ◽  
Atp Binding Cassette

Abstract Background The western corn rootworm, Diabrotica virgifera virgifera, is a pervasive pest of maize in North America and Europe, which has adapted to current pest management strategies. In advance of an assembled and annotated D. v. virgifera genome, we developed transcriptomic resources to use in identifying candidate genes likely to be involved in the evolution of resistance, starting with members of the ATP-binding cassette (ABC) transporter family. Results In this study, 65 putative D. v. virgifera ABC (DvvABC) transporters were identified within a combined transcriptome assembly generated from embryonic, larval, adult male, and adult female RNA-sequence libraries. Phylogenetic analysis placed the deduced amino-acid sequences of the DvvABC transporters into eight subfamilies (A to H). To supplement our sequence data with functional analysis, we identified orthologs of Tribolium castaneum ABC genes which had previously been shown to exhibit overt RNA interference (RNAi) phenotypes. We identified eight such D. v. virgifera genes, and found that they were functionally similar to their T. castaneum counterparts. Interestingly, depletion of DvvABCB_39715 and DvvABCG_3712 transcripts in adult females produced detrimental reproductive and developmental phenotypes, demonstrating the potential of these genes as targets for RNAi-mediated insect control tactics. Conclusions By combining sequence data from four libraries covering three distinct life stages, we have produced a relatively comprehensive de novo transcriptome assembly for D. v. virgifera. Moreover, we have identified 65 members of the ABC transporter family and provided the first insights into the developmental and physiological roles of ABC transporters in this pest species.

scholarly journals ATP-Binding Cassette (ABC) Transporter Genes Involved in Pyrethroid Resistance in the Malaria Vector Anopheles sinensis: Genome-Wide Identification, Characteristics, Phylogenetics, and Expression Profile

2019 ◽  
Vol 20 (6) ◽  
pp. 1409 ◽  
Author(s):  
Qiyi He ◽  
Zhentian Yan ◽  
Fengling Si ◽  
Yong Zhou ◽  
Wenbo Fu ◽  
...  
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Pyrethroid Resistance ◽  
Expression Profiles ◽  
Anopheles Sinensis ◽  
Atp Binding ◽  
Rna Seq ◽  
Transporter Family ◽  
Abc Transporter Genes ◽  
Atp Binding Cassette

background: The ATP-binding cassette (ABC) transporters family is one of the largest families of membrane proteins existing in all living organisms. Pyrethroid resistance has become the largest unique obstacle for mosquito control worldwide. ABC transporters are thought to be associated with pyrethroid resistance in some agricultural pests, but little information is known for mosquitoes. Herein, we investigated the diversity, location, characteristics, phylogenetics, and evolution of ABC transporter family of genes in the Anopheles sinensis genome, and identified the ABC transporter genes associated with pyrethroid resistance through expression profiles using RNA-seq and qPCR. Results: 61 ABC transporter genes are identified and divided into eight subfamilies (ABCA-H), located on 22 different scaffolds. Phylogenetic and evolution analyses with ABC transporters of A. gambiae, Drosophila melanogaster, and Homo sapiens suggest that the ABCD, ABCG, and ABCH subfamilies are monophyly, and that the ABCC and ABCG subfamilies have experienced a gene duplication event. Both RNA-seq and qPCR analyses show that the AsABCG28 gene is uniquely significantly upregulated gene in all three field pyrethroid-resistant populations (Anhui, Chongqing, and Yunnan provinces) in comparison with a laboratory-susceptible strain from Jiangsu province. The AsABCG28 is significantly upregulated at 12-h and 24-h after deltamethrin exposure in three-day-old female adults. Conclusion: This study provides the information frame for ABC transporter subfamily of genes, and lays an important basis for the better understanding and further research of ABC transporter function in insecticide toxification. The AsABCG28 gene is associated with pyrethroid detoxification, and it functions at later period in the detoxification process for xenobiotics transportation.

scholarly journals Structural and functional insights into the Diabrotica virgifera virgifera ATP-binding cassette transporter gene family

2019 ◽  
Author(s):  
Folukemi Adedipe ◽  
Nathaniel Grubbs ◽  
Brad Coates ◽  
Brian Wiegmman ◽  
Marce Lorenzen
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Transcriptome Assembly ◽  
Diabrotica Virgifera Virgifera ◽  
Egg Laying ◽  
Atp Binding ◽  
Pest Species ◽  
Diabrotica Virgifera ◽  
Transporter Family ◽  
Atp Binding Cassette

Abstract Background The western corn rootworm, Diabrotica virgifera virgifera , is a pervasive pest of cultivated maize in North America and Europe, which has adapted to survive exposure to multiple insecticidal agents. Due to their role in insecticide transport, we sought to identify members of the ATP-binding cassette (ABC) transporter family in D. v. virgifera using a transcriptomics approach.Results In this study, 65 putative D. v. virgifera ABC ( Dvv ABC) transporters were identified within a combined transcriptome assembly generated from embryonic, larval, adult male, and adult female RNA-sequence libraries. Phylogenetic analysis placed the deduced amino-acid sequences of the Dvv ABC transporters into eight subfamilies (A to H). Of these, eight shared structural and functional conservation with Tribolium castaneum ABC transporter orthologs known to exhibit overt RNA interference (RNAi) knockdown phenotypes. Interestingly, depletion of DvvABCB_19147 and DvvABCG_3712 transcripts in adult females produced detrimental reproductive and developmental phenotypes (egg-laying or -hatching defects), demonstrating the potential of these genes as targets for RNAi-mediated insect control tactics.Conclusions By combining sequence data from four libraries covering three distinct life stages, we have produced a relatively comprehensive de novo transcriptome assembly for D. v. virgifera . Moreover, we have identified 65 members of the ABC transporter family, and provided the first insights into the developmental and physiological roles of ABC transporters in this pest species.

scholarly journals Function and Role of ATP-Binding Cassette Transporters as Receptors for 3D-Cry Toxins

Toxins ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 124 ◽  
Author(s):  
Ryoichi Sato ◽  
Satomi Adegawa ◽  
Xiaoyi Li ◽  
Shiho Tanaka ◽  
Haruka Endo
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Cultured Cells ◽  
Cell Swelling ◽  
Cry Toxins ◽  
Transporter Family ◽  
Pore Opening ◽  
Synergistic Relationship ◽  
High Binding Affinity

When ABC transporter family C2 (ABCC2) and ABC transporter family B1 (ABCB1) were heterologously expressed in non-susceptible cultured cells, the cells swelled in response to Cry1A and Cry3 toxins, respectively. Consistent with the notion that 3D-Cry toxins form cation-permeable pores, Bombyx mori ABCC2 (BmABCC2) facilitated cation-permeable pore formation by Cry1A when expressed in Xenopus oocytes. Furthermore, BmABCC2 had a high binding affinity (KD) to Cry1Aa of 3.1 × 10−10 M. These findings suggest that ABC transporters, including ABCC2 and ABCB1, are functional receptors for 3D-Cry toxins. In addition, the Cry2 toxins most distant from Cry1A toxins on the phylogenetic tree used ABC transporter A2 as a receptor. These data suggest that 3D-Cry toxins use ABC transporters as receptors. In terms of inducing cell swelling, ABCC2 has greater activity than cadherin-like receptor. The pore opening of ABC transporters was hypothesized to be linked to their receptor function, but this was repudiated by experiments using mutants deficient in export activity. The synergistic relationship between ABCC2 and cadherin-like receptor explains their ability to cause resistance in one species of insect.

CFTR Is a Conductance Regulator as well as a Chloride Channel

1999 ◽  
Vol 79 (1) ◽  
pp. S145-S166 ◽  
Author(s):  
ERIK M. SCHWIEBERT ◽  
DALE J. BENOS ◽  
MARIE E. EGAN ◽  
M. JACKSON STUTTS ◽  
WILLIAM B. GUGGINO
Keyword(s):  
Cystic Fibrosis ◽  
Abc Transporter ◽  
Abc Transporters ◽  
Chloride Channel ◽  
Transmembrane Conductance Regulator ◽  
Cellular Functions ◽  
Small Peptides ◽  
Transmembrane Conductance ◽  
Transporter Family ◽  
Ion Channel Proteins

Schwiebert, Erik M., Dale J. Benos, Marie E. Egan, M. Jackson Stutts, and William B. Guggino. CFTR Is a Conductance Regulator as well as a Chloride Channel. Physiol. Rev. 79, Suppl.: S145–S166, 1999. — Cystic fibrosis transmembrane conductance regulator (CFTR) is a member of the ATP-binding cassette (ABC) transporter gene family. Although CFTR has the structure of a transporter that transports substrates across the membrane in a nonconductive manner, CFTR also has the intrinsic ability to conduct Cl− at much higher rates, a function unique to CFTR among this family of ABC transporters. Because Cl− transport was shown to be lost in cystic fibrosis (CF) epithelia long before the cloning of the CF gene and CFTR, CFTR Cl− channel function was considered to be paramount. Another equally valid perspective of CFTR, however, derives from its membership in a family of transporters that transports a multitude of different substances from chemotherapeutic drugs, to amino acids, to glutathione conjugates, to small peptides in a nonconductive manner. Moreover, at least two members of this ABC transporter family ( mdr-1, SUR) can regulate other ion channels in the membrane. More simply, ABC transporters can regulate somehow the function of other cellular proteins or cellular functions. This review focuses on a plethora of studies showing that CFTR also regulates other ion channel proteins. It is the hope of the authors that the reader will take with him or her the message that CFTR is a conductance regulator as well as a Cl− channel.

scholarly journals ABC Transporters Involved in Export of Cell Surface Glycoconjugates

2010 ◽  
Vol 74 (3) ◽  
pp. 341-362 ◽  
Author(s):  
Leslie Cuthbertson ◽  
Veronica Kos ◽  
Chris Whitfield
Keyword(s):  
Cell Surface ◽  
Abc Transporter ◽  
Abc Transporters ◽  
Nucleotide Binding ◽  
Binding Domain ◽  
Nucleotide Binding Domain ◽  
Polysaccharide Biosynthesis ◽  
O Antigens ◽  
And Function ◽  
Substrate Binding Domain

SUMMARY Complex glycoconjugates play critical roles in the biology of microorganisms. Despite the remarkable diversity in glycan structures and the bacteria that produce them, conserved themes are evident in the biosynthesis-export pathways. One of the primary pathways involves representatives of the ATP-binding cassette (ABC) transporter superfamily. These proteins are responsible for the export of a wide variety of cell surface oligo- and polysaccharides in both Gram-positive and Gram-negative bacteria. Recent investigations of the structure and function of ABC transporters involved in the export of lipopolysaccharide O antigens have revealed two fundamentally different strategies for coupling glycan polymerization to export. These mechanisms are distinguished by the presence (or absence) of characteristic nonreducing terminal modifications on the export substrates, which serve as chain termination and/or export signals, and by the presence (or absence) of a discrete substrate-binding domain in the nucleotide-binding domain polypeptide of the ABC transporter. A bioinformatic survey examining ABC exporters from known oligo- and polysaccharide biosynthesis loci identifies conserved nucleotide-binding domain protein families that correlate well with themes in the structures and assembly of glycans. The familial relationships among the ABC exporters generate hypotheses concerning the biosynthesis of structurally diverse oligo- and polysaccharides, which play important roles in the biology of bacteria with different lifestyles.

scholarly journals Gene expression analysis of ABC transporter family in breast tumors: relationship with chemotherapy effect and disease prognosis

2020 ◽  
Vol 7 (2) ◽  
pp. 29-38
Author(s):  
M. M. Tsyganov ◽  
M. K. Ibragimova ◽  
A. M. Pevzner ◽  
K. A. Gaptulbarova ◽  
E. Yu. Garbukov ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Abc Transporter ◽  
Abc Transporters ◽  
Survival Rates ◽  
Prognostic Significance ◽  
Rank Test ◽  
Transporter Family ◽  
High Gene Expression ◽  
Before And After

Background. One of the main reasons of the ineffectiveness of chemotherapy is still considered to be the formation of the multidrug resistance phenotype of the tumor due to the expression of energy-dependent proteins of ABC transporters. Our previous studies for some ABC genes have established that the expression of these genes correlates with the effectiveness of neoadjuvant chemotherapy (NAC). Some of the clinical studies indicate that ABC transporters can influence not only the formation of chemoresistance in the tumor, but also the progression, invasion and metastasis of the tumor node. Objective: to evaluate the level of transcription of all 49 known ABC genes in a breast tumor before and after treatment and their prognostic significance. Materials and methods. The study included 31 patients with a diagnosis of stage IIA – IIIB breast cancer. RNA was isolated from paired samples of tumor tissue before and after NAC. A microarray study of all tumor samples was performed on ClariomТМ S Assay, human microarrays. Using microarray studies, the expression of 49 genes of the ABC transporter family was studied. Analysis of the microchip data was carried out using the program Transcriptome Analysis Console (TAC) software 4.0. Results. It was found that changes in the expression (increase/decrease during NAC) of the ABCA5, ABCA7, ABCB1, ABCB4, ABCB11, ABCC1, ABCC10, ABCC11, ABCG1, ABCG2, ABCG4, ABCG5, ABCG8 genes are statistically significantly associated with the response to NAC. In addition, the prognostic significance of ABCB1 and ABCB4 gene expression was established. Survival analysis showed that 5-year survival rates in patients with high gene expression of ABCB1 and ABCB4 are lower compared to patients with low expression of these genes (log-rank-test p = 0.001 and 0.04 respectively). Conclusion. Data were obtained on the relationship of gene expression of the ABC transporter family with the effect of NAC in patients with breast cancer and the outcome of the disease. The prognostic potential of the ABCB1 and ABCB4 genes in patients with breast cancer has been established.

scholarly journals Structural and functional insights into the Diabrotica virgifera virgifera ATP-binding cassette transporter gene family

2019 ◽  
Author(s):  
Folukemi Adedipe ◽  
Nathaniel Grubbs ◽  
Brad Coates ◽  
Brian Wiegmman ◽  
Marce Lorenzen
Keyword(s):  
Abc Transporter ◽  
Abc Transporters ◽  
Transcriptome Assembly ◽  
Diabrotica Virgifera Virgifera ◽  
Egg Laying ◽  
Atp Binding ◽  
Pest Species ◽  
Diabrotica Virgifera ◽  
Transporter Family ◽  
Atp Binding Cassette

Abstract Background The western corn rootworm, Diabrotica virgifera virgifera , is a pervasive pest of cultivated maize in North America and Europe, which has adapted to survive exposure to multiple insecticidal agents. Due to their role in insecticide transport, we sought to identify members of the ATP-binding cassette (ABC) transporter family in D. v. virgifera using a transcriptomics approach.Results In this study, 65 putative D. v. virgifera ABC ( Dvv ABC) transporters were identified within a combined transcriptome assembly generated from embryonic, larval, adult male, and adult female RNA-sequence libraries. Phylogenetic analysis placed the deduced amino-acid sequences of the Dvv ABC transporters into eight subfamilies (A to H). Of these, eight shared structural and functional conservation with Tribolium castaneum ABC transporter orthologs known to exhibit overt RNA interference (RNAi) knockdown phenotypes. Interestingly, depletion of DvvABCB_19147 and DvvABCG_3712 transcripts in adult females produced detrimental reproductive and developmental phenotypes (egg-laying or -hatching defects), demonstrating the potential of these genes as targets for RNAi-mediated insect control tactics.Conclusions By combining sequence data from four libraries covering three distinct life stages, we have produced a relatively comprehensive de novo transcriptome assembly for D. v. virgifera . Moreover, we have identified 65 members of the ABC transporter family, and provided the first insights into the developmental and physiological roles of ABC transporters in this pest species.

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