Survey of cellular factors modulating the HIV 1 integration complex activity using a unique protein screening system in vitro

... ubiquitin ligases in the degradation of HIV- 1 IN 14 1. 6 HIV- 1 PIC as a better target of study than recombinant IN 17 1. 6 .1 Cellular components and modulators of the pre -integration nucleoprotein complex ... Structural domain of HIV- 1 IN IN contains 288 amino acid residues and has three protein domains The NTD facilitates protein dimerization, CCD is involved in the catalysis of integration, and CTD has ... mechanism in the enhancement of HIV- 1 integration VI LIST OF TABLES Table 1. 1 List of cellular cofactors that interact with IN to 12 modulate HIV- 1 replication processes in the early phase Table 1. 2...

Báo cáo y học: " siRNA and shRNA screens advance key understanding of host factors required for HIV-1 replication" potx

... reports of siRNA/ shRNA screens for HIV-1 replication cofactors are therefore not the beginning of the end, but the end of the beginning of a new era in the search for host factors required for HIV-1 ... exhaustive The goal of siRNA and shRNA screens is to identify all possible HIV-1 cofactors For productive replication to occur, HIV-1 has to switch on and off many cellular pathways Plausibly, at different ... suggested to account for the identification of the many different HIV-1 cofactors in the screens [1] Figure for HIV-1 replication cofactors A comparison between siRNA- and shRNA- based screens A comparison...

Báo cáo y học: " Modulation of microtubule assembly by the HIV-1 Tat protein is strongly dependent on zinc binding to Tat" pptx

... Since apoptosis mediated by Tat partly relies on the interaction of Tat with tubulin [14-17], we hypothesized that zinc binding might play a role in the modulation by Tat of the microtubule dynamics ... 5:62 Consequently, holo -Tat likely constitutes the active form of Tat in the cell cytoplasm By altering the microtubule dynamics, holo -Tat may then lead to a release of the proapoptotic Bim protein, ... non -assembly conditions Zinc binding to Tat promotes discrete Tat- tubulin complexes Zinc binding to Tat promotes discrete Tat- tubulin complexes under non -assembly conditions A Characterization...

Tài liệu Báo cáo khoa học: Different mechanisms for cellular internalization of the HIV-1 Tat-derived cell penetrating peptide and recombinant proteins fused to Tat docx

... FACS analysis of the internalization of the full-length Tat protein construct and the Tat CPP Differences in the mechanisms of internalization between the Tat peptide and the Tat fused protein ... at the surface of the Tat- containing recombinant proteins and call other reasons to explain the differences in the mechanism of internalization between the CPP peptide and the Tat- containing proteins ... alter the uptake of the Tat CPP These biochemical evidences con®rmed a pathway for the entry of the Tat peptide unrelated to the HS proteoglycan receptors Internalization of the Tat CPP did not...

Báo cáo hóa học: " Mapping of immunogenic and protein-interacting regions at the surface of the seven-bladed β-propeller domain of the HIV-1 cellular interactor EED" pptx

... our mapping of the MA binding site on the EED linear sequence, since β-strands d were the most exposed β-strands at the periphery of the β-propeller (Fig 2D) Of note, the upper face of the β-propeller ... β-propeller domain of EED and protein-interacting regions Figure Surface representation of the β-propeller domain of EED and protein-interacting regions The binding residues of HIV-1 proteins are ... Thus, at the early phase of the HIV-1 life cycle, EED might play a role in targeting the regions of proviral DNA integration into the host chromatin At the late steps of the virus replication cycle,...

Báo cáo y học: " Changes in the accessibility of the HIV-1 Integrase C-terminus in the presence of cellular proteins" doc

... added a biotinylable tag at the C-terminus of IN (IN- BAD) and showed that the protein remains fully active in the context of a lentiviral vector The kinetics of viral DNA synthesis and integration ... shown) The presence of LEDGF/p75 in infected cells prevents access to the IN C-terminus We next asked whether the presence of LEDGF/p75 in cells lysates could be linked directly or indirectly to the ... masking of the biotinylated tag in the context of PICs To resolve these issues, we analysed the presence of IN in our samples (the same extract used in SA capture experiment shown in Fig 2B) by...

Báo cáo y học: " Intermolecular masking of the HIV-1 Rev NLS by the cellular protein HIC: Novel insights into the regulation of Rev nuclear impo" doc

... necessary and sufficient for Rev nuclear import inhibition by HIC Nuclear imports of GST-YFP -Rev, GST-YFP -Rev N1, GST-YFP -Rev N2 and GST-YFP-RevNLS were examined using in vitro nuclear import assays ... pathways or the physical obstruction of the NPC since M9 or Rev nuclear import mediated by transportin remained unaffected by HIC The molecular recognition of NLSs by import receptors in the cytoplasm ... propose that the intermolecular masking of Rev NLS by HIC by which HIC control of Rev nuclear import can contribute to the spatial control of its activity We also show that Rev nuclear import...

Báo cáo y học: "HIV-1 integrase modulates the interaction of the HIV-1 cellular cofactor LEDGF/p75 with chromati" pptx

... [33] The surface of interaction of the IBD with a subset of these proteins partially overlaps with that of integrase (reviewed in [19]) The affinity of IBD for integrase is higher than for some of ... integrase The HIV-1 co-factor activity of LEDGF/p75 involves its simultaneous interaction with the host chromatin and with HIV-1 integrase [1,2] To determine the effect of the ionic strength on the interaction ... (this study and [7]) The enhancing effect of integrase on the chromatin binding strength of LEDGF/p75 could offer a molecular explanation for the conserved HIV-1 cofactor activity of the LEDGF/p75...

Báo cáo y học: " Induction of the HIV-1 Tat co-factor cyclin T1 during monocyte differentiation is required for the regulated expression of a large portion of cellular mRNAs" pptx

... manuscript YW and XQ constructed and characterized the HIV-1 viruses and lentiviral vectors used in the study CS performed the analysis of DNA microarray data and performed the statistical analysis AR ... protocol (Promega), and the products were measured by a luminometer (Turner) Microarray analysis Microarray analysis was performed by the Baylor Microarray Core Facility (Baylor College of Medicine, ... cyclin T1/ PTEFb complex as a cofactor, the virus may assure an optimal cellular environment for maximal virus production Further analysis of the genes regulated by cyclin T1 may lead to the identification...

Báo cáo khoa học: Structural mobility of the monomeric C-terminal domain of the HIV-1 capsid protein pptx

... Ser146 in the numbering of the intact CA) The CACW40A protein is monomeric, and its structure is similar to that of the subunits in the dimeric, non-mutated CAC, but, in the monomeric form, the segment ... responsible, from a thermodynamic point of view, for the binding of the monomeric species of CAC? We have previously discussed the variation in the free energy of binding as a function of the changes ... dimerization domain of the HIV-1 capsid protein Science 278, 849–853 23 Worthylake DK, Wang H, Yoo S, Sundquist WI & Hill CP (1999) Structures of the HIV-1 capsid protein ˚ dimerization domain at...

Báo cáo khoa học: Kinetic study of the HIV)1 DNA 3¢-end processing Single-turnover property of integrase docx

... to the processed DNA product The stability of IN during the time-course of the reaction and the inuence of protein aggregation are discussed Results General features of the 3Â-end processing kinetics ... clear In the case of IN, tight binding of the protein to recessed viral DNA ends is obligatory in the context of the HIV replication cycle: the 3Â -processing of viral DNA ends is only the rst ... 0.06 Taking into account the standard duration of 3Â -processing assays, the absence of turnover in the case of IN originates primarily in the low kcatÂ value The amount of INDNA complex, which did...

Báo cáo Y học: Structural basis for the inhibitory efﬁcacy of efavirenz (DMP-266), MSC194 and PNU142721 towards the HIV-1 RT K103N mutant doc

... with the symmetry of space group C2221 Overall hydrophobic NNRTI interactions to wild-type and K103N mutant RT The interactions of Efavirenz, MSC194 and PNU142721 to wild-type RT and the K103N mutant ... study we present the structural indications for the role of K103 and N103 in drug binding and the structural implications for the inhibitory efficacy of the inhibitors Efavirenz, PNU142721, and MSC194 ... potency The inhibitory efficacy to wild-type RT is within subnanomolar range for all three inhibitors PNU142721 and MSC194 show a 3–10-fold reduction in efficacy for the K103N mutant The effect of the...

Báo cáo khoa học: Alizarine derivatives as new dual inhibitors of the HIV-1 reverse transcriptase-associated DNA polymerase and RNase H activities effective also on the RNase H activity of non-nucleoside resistant reverse transcriptases pot

... which was slightly active on the wild-type RT RNase H function, showed a sixfold increase in the inhibition potency of the RNase H function, although it retained its inhibition potency on the ... AQ derivatives and other RT inhibitors (A) YonetaniTheorell plot of the combination of K-49 and RDS 1643 on the HIV-1 RT polymeraseindependent RNase H activity HIV-1 RT was incubated in the presence ... [25] Therefore, the HIV-1 RT RNase H activity was measured in the presence of increasing concentrations of both K-49 and RDS1643, and was analyzed using the YonetaniTheorell plot (Fig 2) The results...

Báo cáo khoa học: Peptides that bind the HIV-1 integrase and modulate its enzymatic activity – kinetic studies and mode of action pptx

... on the 3Â-end processing step (not shown) The effect of peptides on the strand-transfer reaction When the effects of the various peptides on the strandtransfer step of the IN enzymatic reaction ... group of peptides is attributable to their effect on the turnover number of IN, and suggest a different mode of action from that of the rst group The third group of peptides, which included IN-4 and ... resembles that of INS [56] To convert some of these peptides into efcient antiHIV drugs, it is essential to elucidate their mode of action and their effects on the kinetic parameters of the 3Â-end...

Báo cáo Y học: NMR structure of the HIV-1 regulatory protein Vpr in H2O/triﬂuoroethanol Comparison with the Vpr N-terminal (1–51) and C-terminal (52–96) domains pot

... binding of NCp7 to the C-terminal domain (80–96 )Vpr [8] may expose the N-terminal (1–39) portion of the protein and allow its tight binding to Lys-tRNA synthetase [15] The C-terminal domain (52–96 )Vpr ... when comparing the isolated domains of (1–51 )Vpr [25] and (52–96 )Vpr [26] with the intact Vpr protein The differences in structure are localized to the N-terminal domain The secondary structures ... of the a-helices in the 3D structure of the protein The backbone is represented in blue and the hydrophobic amino acid side chain coloured pink Ó FEBS 2002 NMR Structure of the HIV-1 protein Vpr...

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