The process of integrating the reverse-transcribed HIV-1 DNA into the host chromosomal DNA is catalyzed by the virally encoded enzyme integrase (IN). Integration requires two metal-dependent reactions, 3 end processing and strand transfer. Compounds that contain a diketo acid moiety have been shown to selectively inhibit the strand transfer reaction of IN in vitro and in infected cells and are effective as inhibitors of HIV-1 replication. To characterize the molecular basis of inhibition, we used functional assays and binding assays to evaluate a series of structurally related analogs. These studies focused on investigating the role of the conserved carboxylate and metal binding. We demonstrate that an acidic moiety such as a carboxylate or isosteric heterocycle is not required for binding to the enzyme complex but is essential for inhibition and confers distinct metaldependent properties on the inhibitor. Binding requires divalent metal and resistance is metal dependent with active site mutants displaying resistance only when the enzymes are evaluated in the context of Mg 2؉ . The mechanism of action of these inhibitors is therefore likely a consequence of the interaction between the acid moiety and metal ion(s) in the IN active site, resulting in a functional sequestration of the critical metal cofactor(s). These studies thus have implications for modeling active site inhibitors of IN, designing and evaluating analogs with improved efficacy, and identifying inhibitors of other metal-dependent phosphotransferases.A n essential step in HIV replication is the integration of the reverse-transcribed viral genome into host chromosomal DNA by the virally encoded integrase (IN) protein (1-3). Integration is required for efficient long terminal repeat-driven transcription of the provirus for the production of viral proteins and RNA progeny. IN represents an important chemotherapeutic target, as its inactivation, either by mutagenesis or inhibition, blocks productive infection by HIV-1 (4-7).Integration is carried out in the cell in a series of distinct steps (8-10). First, IN cleaves the two terminal nucleotides from each 3Ј end of the viral DNA. The 3Ј processing reaction is carried out concurrently with or soon after reverse transcription in the cytoplasm. In the second step, strand transfer, IN catalyzes staggered nicking of the target chromosomal DNA and joining of each 3Ј end of the viral DNA to the 5Ј ends of the host DNA. Strand transfer is temporally and spatially separated from 3Ј processing and occurs after transport of the preintegration complex from the cytoplasm into the nucleus.Divalent metals such as Mg 2ϩ or Mn 2ϩ are required for both 3Ј processing and strand transfer and for the assembly of IN onto specific viral donor DNA to form a complex competent to carry out either function (11)(12)(13) (18), L-731,988 and related DKAs inhibit integration and viral replication in cell culture (7). Mutations that confer resistance to DKAs have been identified and map to IN residues adjacent to D64 and E152. The in...

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Terrain Awareness and Warning Systems (TAWS) and Synthetic Vision Systems (SVS) provide pilots with displays of stored geo-spatial data (e.g. terrain, obstacles, and/or features). As comprehensive validation is impractical, these databases typically have no quantifiable level of integrity. This lack of a quantifiable integrity level is one of the constraints that has limited categories: Controlled Flight Into Terrain (CFIT) and Loss of Control in flight (LOC) are focal points of both agency's initiatives. The primary causal factor leading to these CFIT and LOC accidents is loss of situational awareness (SA) by pilots with respect to their spatial location and orientation relative to hazards such as terrain and obstacles. Candidate interventions such as SVS and TAWS seek to improve this aspect of SA.certification and operational approval of TAWSISVS to "advisory-only" systems for civil aviation. Previous work demonstrated the feasibility of using a real-time monitor to bound database integrity by using downward-looking remote sensing technology (i.e. radar altimeters). This paper describes an extension of the integrity monitor concept to incIude a forward-looking sensor to cover additional classes of terrain database faults and to reduce the exposure time associated with integrity threats. An operational concept is presented that combines established feature extraction techniques with a statistical assessment of similarity measures between the sensed and stored features using principles from classical detection theoq. Finally, an implementation is presented that uses existing commercial-off-the-shelf weather radar sensor technology.