Multiparameter single-molecule fluorescence spectroscopy reveals heterogeneity of HIV-1 reverse transcriptase:primer/template complexes.

Abstract:

:By using single-molecule multiparameter fluorescence detection, fluorescence resonance energy transfer experiments, and newly developed data analysis methods, this study demonstrates directly the existence of three structurally distinct forms of reverse transcriptase (RT):nucleic acid complexes in solution. Single-molecule multiparameter fluorescence detection also provides first information on the structure of a complex not observed by x-ray crystallography. This species did not incorporate nucleotides and is structurally distinct from the other two observed species. We determined that the nucleic acid substrate is bound at a site far removed from the nucleic acid-binding tract observed by crystallography. In contrast, the other two states are identified as being similar to the x-ray crystal structure and represent distinct enzymatically productive stages in DNA polymerization. These species differ by only a 5-A shift in the position of the nucleic acid. Addition of nucleoside triphosphate or of inorganic pyrophosphate allowed us to assign them as the educt and product state in the polymerization reaction cycle; i.e., the educt state is a complex in which the nucleic acid is positioned to allow nucleotide incorporation. The second RT:nucleic acid complex is the product state, which is formed immediately after nucleotide incorporation, but before RT translates to the next nucleotide.

authors

Rothwell PJ,Berger S,Kensch O,Felekyan S,Antonik M,Wöhrl BM,Restle T,Goody RS,Seidel CA

doi

10.1073/pnas.0434003100

keywords:

subject

Has Abstract

pub_date

2003-02-18 00:00:00

pages

1655-60

issue

4

eissn

0027-8424

issn

1091-6490

pii

0434003100

journal_volume

100

pub_type

杂志文章
  • Mapping multisensory parietal face and body areas in humans.

    abstract::Detection and avoidance of impending obstacles is crucial to preventing head and body injuries in daily life. To safely avoid obstacles, locations of objects approaching the body surface are usually detected via the visual system and then used by the motor system to guide defensive movements. Mediating between visual ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1207946109

    authors: Huang RS,Chen CF,Tran AT,Holstein KL,Sereno MI

    更新日期:2012-10-30 00:00:00

  • Neural correlates of behavior in the moth Manduca sexta in response to complex odors.

    abstract::With Manduca sexta as a model system, we analyzed how natural odor mixtures that are most effective in eliciting flight and foraging behaviors are encoded in the primary olfactory center in the brain, the antennal lobe. We used gas chromatography coupled with multiunit neural-ensemble recording to identify key odorant...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0910592106

    authors: Riffell JA,Lei H,Hildebrand JG

    更新日期:2009-11-17 00:00:00

  • Computing trisections of 4-manifolds.

    abstract::We describe an algorithm to compute trisections of orientable four-manifolds using arbitrary triangulations as input. This results in explicit complexity bounds for the trisection genus of a 4-manifold in terms of the number of pentachora (4-simplices) in a triangulation. ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1717173115

    authors: Bell M,Hass J,Rubinstein JH,Tillmann S

    更新日期:2018-10-23 00:00:00

  • CXCR4 utilization is sufficient to trigger CD4+ T cell depletion in HIV-1-infected human lymphoid tissue.

    abstract::The human chemokine receptors CCR5 and CXCR4 have emerged as the predominant cofactors, along with CD4, for cellular entry of HIV-1 in vivo whereas the contribution of other chemokine receptors to HIV disease has not been yet determined. CCR5-specific (R5) viruses predominate during primary HIV-1 infection whereas vir...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.2.663

    authors: Penn ML,Grivel JC,Schramm B,Goldsmith MA,Margolis L

    更新日期:1999-01-19 00:00:00

  • Crystal structure of the stimulatory complex of GTP cyclohydrolase I and its feedback regulatory protein GFRP.

    abstract::In the presence of phenylalanine, GTP cyclohydrolase I feedback regulatory protein (GFRP) forms a stimulatory 360-kDa complex with GTP cyclohydrolase I (GTPCHI), which is the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin. The crystal structure of the stimulatory complex reveals that the GTPCHI decame...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.022646999

    authors: Maita N,Okada K,Hatakeyama K,Hakoshima T

    更新日期:2002-02-05 00:00:00

  • At least six forms of extremely homologous cytochromes P-450 in rat liver are encoded at two closely linked genetic loci.

    abstract::A subpopulation of phenobarbital-induced cytochrome P-450 in rat liver has been shown to consist of four closely related forms of the enzyme that appeared to be strain-related. In the present study, polypeptides composing this family were analyzed by two-dimensional electrophoresis of hepatic microsomes from 64 indivi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.21.6542

    authors: Rampersaud A,Walz FG Jr

    更新日期:1983-11-01 00:00:00

  • Structure of RavA MoxR AAA+ protein reveals the design principles of a molecular cage modulating the inducible lysine decarboxylase activity.

    abstract::The MoxR family of AAA+ ATPases is widespread throughout bacteria and archaea but remains poorly characterized. We recently found that the Escherichia coli MoxR protein, RavA (Regulatory ATPase variant A), tightly interacts with the inducible lysine decarboxylase, LdcI/CadA, to form a unique cage-like structure. Here,...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1009092107

    authors: El Bakkouri M,Gutsche I,Kanjee U,Zhao B,Yu M,Goret G,Schoehn G,Burmeister WP,Houry WA

    更新日期:2010-12-28 00:00:00

  • Model for eukaryotic tail-anchored protein binding based on the structure of Get3.

    abstract::The Get3 ATPase directs the delivery of tail-anchored (TA) proteins to the endoplasmic reticulum (ER). TA-proteins are characterized by having a single transmembrane helix (TM) at their extreme C terminus and include many essential proteins, such as SNAREs, apoptosis factors, and protein translocation components. Thes...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0907522106

    authors: Suloway CJ,Chartron JW,Zaslaver M,Clemons WM Jr

    更新日期:2009-09-01 00:00:00

  • Functional traits predict relationship between plant abundance dynamic and long-term climate warming.

    abstract::Predicting climate change impact on ecosystem structure and services is one of the most important challenges in ecology. Until now, plant species response to climate change has been described at the level of fixed plant functional types, an approach limited by its inflexibility as there is much interspecific functiona...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1310700110

    authors: Soudzilovskaia NA,Elumeeva TG,Onipchenko VG,Shidakov II,Salpagarova FS,Khubiev AB,Tekeev DK,Cornelissen JH

    更新日期:2013-11-05 00:00:00

  • Affinity of IDPs to their targets is modulated by ion-specific changes in kinetics and residual structure.

    abstract::Intrinsically disordered proteins (IDPs) are characterized by a lack of defined structure. Instead, they populate ensembles of rapidly interconverting conformations with marginal structural stabilities. Changes in solution conditions such as temperature and crowding agents consequently affect IDPs more than their fold...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1705105114

    authors: Wicky BIM,Shammas SL,Clarke J

    更新日期:2017-09-12 00:00:00

  • Androgen-induced proliferative quiescence in prostate cancer cells: the role of AS3 as its mediator.

    abstract::In the prostate gland of adult mammals, most epithelial cells are in a state of proliferative quiescence. Androgens regulate this effect by inducing cell cycle arrest in the G(0)/G(1) phase. Potential mediators of this androgen-induced proliferative shutoff were identified by means of subtracted cDNA libraries. The ex...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.97.18.10185

    authors: Geck P,Maffini MV,Szelei J,Sonnenschein C,Soto AM

    更新日期:2000-08-29 00:00:00

  • Synthetic peptide with cell attachment activity of fibronectin.

    abstract::Four synthetic peptides that together constitute the cell attachment domain of fibronectin [Pierschbacher, M.D., Ruoslahti, E., Sundelin, J., Lind, P. & Peterson, P. (1982) J. Biol. Chem. 257, 9593-9597] were constructed and tested for their ability to induce cell attachment and spreading. One of these peptides, consi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.5.1224

    authors: Pierschbacher M,Hayman EG,Ruoslahti E

    更新日期:1983-03-01 00:00:00

  • The karrikin signaling regulator SMAX1 controls Lotus japonicus root and root hair development by suppressing ethylene biosynthesis.

    abstract::An evolutionarily ancient plant hormone receptor complex comprising the α/β-fold hydrolase receptor KARRIKIN INSENSITIVE 2 (KAI2) and the F-box protein MORE AXILLARY GROWTH 2 (MAX2) mediates a range of developmental responses to smoke-derived butenolides called karrikins (KARs) and to yet elusive endogenous KAI2 ligan...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.2006111117

    authors: Carbonnel S,Das D,Varshney K,Kolodziej MC,Villaécija-Aguilar JA,Gutjahr C

    更新日期:2020-09-01 00:00:00

  • Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria.

    abstract::The unique ability of bovine papillomavirus (BPV-1) DNA to replicate as a stable, multicopy plasmid in transformed mouse cells has led to its utilization as a eukaryotic cloning vector. One limitation of the system has been the marked reduction in transformation efficiency when BPV-1 DNA or the subgenomic transforming...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.23.7147

    authors: Sarver N,Byrne JC,Howley PM

    更新日期:1982-12-01 00:00:00

  • Glucagon receptor activates extracellular signal-regulated protein kinase 1/2 via cAMP-dependent protein kinase.

    abstract::We prepared a stable cell line expressing the glucagon receptor to characterize the effect of G(s)-coupled receptor stimulation on extracellular signal-regulated protein kinase 1/2 (ERK1/2) activity. Glucagon treatment of the cell line caused a dose-dependent increase in cAMP concentration, activation of cAMP-dependen...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.131200398

    authors: Jiang Y,Cypess AM,Muse ED,Wu CR,Unson CG,Merrifield RB,Sakmar TP

    更新日期:2001-08-28 00:00:00

  • Human type I procollagen genes are located on different chromosomes.

    abstract::A recombinant plasmid containing sequences complementary to human pro-alpha l(I) collagen mRNA was used for the chromosomal assignment of the pro-alpha l(I) collagen gene. Restriction endonuclease analysis of DNA from mouse-human and Chinese hamster-human somatic cell hybrids revealed cosegregation with human chromoso...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.21.6627

    authors: Huerre C,Junien C,Weil D,Chu ML,Morabito M,Van Cong N,Myers JC,Foubert C,Gross MS,Prockop DJ,Boué A,Kaplan JC,de la Chapelle A,Ramirez F

    更新日期:1982-11-01 00:00:00

  • Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia.

    abstract::Micro-RNAs (miR genes) are a large family of highly conserved noncoding genes thought to be involved in temporal and tissue-specific gene regulation. MiRs are transcribed as short hairpin precursors ( approximately 70 nt) and are processed into active 21- to 22-nt RNAs by Dicer, a ribonuclease that recognizes target m...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.242606799

    authors: Calin GA,Dumitru CD,Shimizu M,Bichi R,Zupo S,Noch E,Aldler H,Rattan S,Keating M,Rai K,Rassenti L,Kipps T,Negrini M,Bullrich F,Croce CM

    更新日期:2002-11-26 00:00:00

  • The isolation of simian virus 40 variants with specifically altered genomes.

    abstract::Serial passage of simian virus 40 (SV40) at high multiplicity of infection leads to the emergence of variants with deleted, substituted, and/or duplicated DNA. Individual variants have been cloned by selective complementation with temperature sensitive SV40 mutants, or nonselectively by coinfection of cells with wild-...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.71.3.942

    authors: Brockman WW,Nathans D

    更新日期:1974-03-01 00:00:00

  • Biochemical evidence for the autophosphorylation and transphosphorylation of transforming growth factor beta receptor kinases.

    abstract::Transforming growth factor beta (TGF-beta) signals through a receptor complex containing the type I (TGF-beta RI) and type II (TGF-beta RII) receptors. We describe here biochemical studies on early events in the TGF-beta signaling pathways. TGF-beta RII is highly phosphorylated when expressed alone in COS-1 cells; its...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.92.5.1565

    authors: Chen F,Weinberg RA

    更新日期:1995-02-28 00:00:00

  • Physical studies of denatured tRNA2Glu from Escherichia coli.

    abstract::We have examined the 270 MHz nuclear magnetic resonance spectrum and relaxation kinetic behavior of tRNA2Glu (E. coli) in the absence of Mg++, a condition which produces an inactive form of this tRNA. The results show that the denatured form has about five fewer proton resonances in the region from -12 to -15 ppm. Rel...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.73.7.2216

    authors: Bina-Stein M,Crothers DM,Hilbers CW,Shulman RG

    更新日期:1976-07-01 00:00:00

  • The point of no return in vetoing self-initiated movements.

    abstract::In humans, spontaneous movements are often preceded by early brain signals. One such signal is the readiness potential (RP) that gradually arises within the last second preceding a movement. An important question is whether people are able to cancel movements after the elicitation of such RPs, and if so until which po...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1513569112

    authors: Schultze-Kraft M,Birman D,Rusconi M,Allefeld C,Görgen K,Dähne S,Blankertz B,Haynes JD

    更新日期:2016-01-26 00:00:00

  • Regulation of human monocyte adherence by granulocyte-macrophage colony-stimulating factor.

    abstract::Recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) was found to increase the adherence of purified peripheral blood monocytes to plastic surfaces and to monolayers of human umbilical vein endothelial cells. With plastic surfaces as a model 9-hr culture with GM-CSF was necessary for enhancement...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.86.18.7169

    authors: Gamble JR,Elliott MJ,Jaipargas E,Lopez AF,Vadas MA

    更新日期:1989-09-01 00:00:00

  • Speaker gaze increases information coupling between infant and adult brains.

    abstract::When infants and adults communicate, they exchange social signals of availability and communicative intention such as eye gaze. Previous research indicates that when communication is successful, close temporal dependencies arise between adult speakers' and listeners' neural activity. However, it is not known whether s...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1702493114

    authors: Leong V,Byrne E,Clackson K,Georgieva S,Lam S,Wass S

    更新日期:2017-12-12 00:00:00

  • Killer whale presence drives bowhead whale selection for sea ice in Arctic seascapes of fear.

    abstract::The effects of predator intimidation on habitat use and behavior of prey species are rarely quantified for large marine vertebrates over ecologically relevant scales. Using state space movement models followed by a series of step selection functions, we analyzed movement data of concurrently tracked prey, bowhead whal...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1911761117

    authors: Matthews CJD,Breed GA,LeBlanc B,Ferguson SH

    更新日期:2020-03-24 00:00:00

  • Defective glycerol metabolism in aquaporin 9 (AQP9) knockout mice.

    abstract::Aquaporin-9 (AQP9) is an aquaglyceroporin membrane channel shown biophysically to conduct water, glycerol, and other small solutes. Because the physiological role/s of AQP9 remain undefined and the expression sites of AQP9 remain incomplete and conflicting, we generated AQP9 knockout mice. In the absence of physiologi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0610894104

    authors: Rojek AM,Skowronski MT,Füchtbauer EM,Füchtbauer AC,Fenton RA,Agre P,Frøkiaer J,Nielsen S

    更新日期:2007-02-27 00:00:00

  • Dephosphorylation of the nuclear factor of activated T cells (NFAT) transcription factor is regulated by an RNA-protein scaffold complex.

    abstract::Nuclear factor of activated T cells (NFAT) proteins are Ca(2+)-regulated transcription factors that control gene expression in many cell types. NFAT proteins are heavily phosphorylated and reside in the cytoplasm of resting cells; when cells are stimulated by a rise in intracellular Ca(2+), NFAT proteins are dephospho...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1019711108

    authors: Sharma S,Findlay GM,Bandukwala HS,Oberdoerffer S,Baust B,Li Z,Schmidt V,Hogan PG,Sacks DB,Rao A

    更新日期:2011-07-12 00:00:00

  • Release of protein A from the cell wall of Staphylococcus aureus.

    abstract::Staphylococcal protein A (SpA) is anchored to the cell wall envelope of Staphylococcus aureus by sortase A, which links the threonyl (T) of its C-terminal LPXTG motif to peptidoglycan cross-bridges (i.e., Gly5). SpA binds the Fcγ domains of IgG and protects staphylococci from opsonophagocytic clearance. Moreover, SpA ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1317181111

    authors: Becker S,Frankel MB,Schneewind O,Missiakas D

    更新日期:2014-01-28 00:00:00

  • Chromosomal mapping of the mink cell focus-inducing and xenotropic env gene family in the mouse.

    abstract::Chromosomal locations of members of the xenotropic-related env gene family in the mouse genome have been determined. Endonuclease restriction site polymorphisms detected by molecular hybridization were used to study the inheritance of mink cell-focus inducing and xenotropic env gene-related sequences in recombinant in...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.20.6298

    authors: Blatt C,Mileham K,Haas M,Nesbitt MN,Harper ME,Simon MI

    更新日期:1983-10-01 00:00:00

  • Receptor- and heparin-binding domains of basic fibroblast growth factor.

    abstract::Two functional domains in the primary structure of basic fibroblast growth factor (FGF) have been identified on the basis of their ability to interact with the FGF receptor, bind radiolabeled heparin, and modulate the cellular response to FGF. Peptides derived from these two functional domains can act as partial agoni...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.85.7.2324

    authors: Baird A,Schubert D,Ling N,Guillemin R

    更新日期:1988-04-01 00:00:00

  • Cleavage of Rous sarcoma viral polypeptide precursor into internal structural proteins in vitro involves viral protein p15.

    abstract::The polypeptide precursor pr76 to the internal viral group specific (gs) antigen proteins of Rous sarcoma virus, synthesized in a cell-free system of ascites cells, has been processed in vitro into the viral proteins by purified viral protein p15 as well as by disrupted Rous sarcoma virus. Disrupted Rauscher murine le...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.74.3.911

    authors: von der Helm K

    更新日期:1977-03-01 00:00:00