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Topic: Integrase

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	Early Research on Integrase Inhibitors: 2 classes of compounds identified
		Integrase functions in an as-yet biochemically undefined complex of viral and cellular proteins referred to as the preintegration complex (Farnet and Haseltine, 1991 and Brown et al., 1987).
		Integrase was added to 20 ul of resin in a 4:1 molar ratio of integrase to DNA (1.92 nmol per sample) and incubated for 30 min.
		Integrase is a 288 amino acid protein with a codon usage that is skewed against high-level expression in E. coli.
	www.natap.org /2004/HIV/101804_04.htm (5133 words)

	AEGiS-GMHC: HIV Integrase: A New Therapeutic Target
		HIV integrase is the virus's third enzyme (the other two are reverse transcriptase and protease).
		Integrase as a therapeutic target does have several possible pitfalls.
		Conversely, however, the intracellular levels of integrase may be extremely small, and this scarcity works in favor of an integrase inhibitor.
	www.aegis.com /pubs/gmhc/1995/gm090202.html (883 words)

	The Body: Bottom of the Ninth: With Two Down And Integrase On Deck, AIDS Research Team Hopes For a Hit--Or Will It Be ...
		Blocking integrase is widely considered a viable target for potential anti-HIV drugs, since recombinant (genetically altered) HIV virions that lack the integrase gene are incapable of infecting and killing cells.
		Integrase inhibitors, on the other hand, have only one chance to inhibit the enzyme and once this opportunity is lost the inhibitor becomes ineffective for that cell.
		As integrase represents the third and final HIV native enzyme protein encoded in the HIV genome, exploring it as a therapeutic target should be, and has become, a major research priority.
	www.thebody.com /tag/tagapr2.html (924 words)

	Overexpression and Purification of Integrase
		One of the major reasons for the difficulty in purifying the Integrase protein is that overexpression of the protein in E. coli cells is toxic.
		The binding of Integrase to the DNA is monitored by analyzing the molecular weight of the radioactively-labeled DNA.
		Upon binding of the Integrase protein to the DNA, the radioactively-labeled DNA would move slower through the gel matrix due to the larger size of the DNA-Integrase complex as compared to the unbound DNA, and this is monitored by autoradiography following acrylamide gel electrophoresis.
	www.sci.sdsu.edu /classes/chemistry/chem467l/shubeita/experiment3.htm (1811 words)

	The Body: Anti-HIV Pipeline: Spotlight on Integrase Inhibitors
		Integrase inhibitors seek to block the integrase enzyme from allowing this integration process from happening.
		Also, the integrase enzyme doesn't occur naturally in the body, so blocking it might not cause some of the side effects common to the existing drugs.
		While it has taken longer for integrase inhibitors to make it into human testing than the current classes of drugs, the failure and/or delay of these first two is no reason to write off the entire class.
	www.thebody.com /pinf/jul04/integrase.html (893 words)

	UCI College of Medicine: Dept. of Pathology: Faculty : W. Edward Robinson, Jr., M.D., Ph.D.
		Integrase is absolutely required for HIV replication and has no mammalian homolog.
		Furthermore, integrase is the third of three viral enzymes and the only enzyme for which no inhibitors are used to treat patients.
		Human immunodeficiency virus type-1 integrase containing a glycine to serine mutation at position 140 is attenuated for catalysis and resistant to integrase inhibitors.
	www.ucihs.uci.edu /com/pathology/faculty/erobinson.html (763 words)

	Early Research on Integrase Inhibitors: 2 Classes of Compounds Identified (Site not responding. Last check: 2007-10-08)
		It has been proposed that this is due to a significant conformational change that occurs on assembly of the integrase-viral DNA complex (Espeseth et al., 2000).
		The assay only performs the strand transfer integration step, because a preformed complex of integrase and donor DNA is used and the donor DNA lacks the two 3’-terminal nucleotides that are removed by integrase in vivo.
		Removal of unbound integrase from the donor-integrase complex was essential to prevent nonspecific binding of integrase to the target DNA, blocking integration.
	www.hivdent.org /drugs1/drugERII102004.htm (5139 words)

	W0283: Crystallization of a Feline Immunodeficiency Virus Integrase Fusion Protein (Site not responding. Last check: 2007-10-08)
		Integrase catalyzes integration of the cDNA copy of the viral genome into the host chromosome, a necessary step in the retroviral life cycle.
		Integrase processes the U5 and U3 termini of the viral DNA by cleaving off two terminal nucleotides, and leaving a recessed 3'-hydroxyl on both ends.
		The integrase proteins of human and feline immunodeficiency viruses (HIV and FIV) share about 86% similarity and 37% identity and have similar biochemical activities and sub-domain organization.
	www.hwi.buffalo.edu /ACA/ACA98/abstracts/text/W0283.html (248 words)

	Probing Interactions between Viral DNA and Human Immunodeficiency Virus Type 1 Integrase Using Dinucleotides -- ...
		Retroviral integrases are essential for viral replication and represent an attractive chemotherapeutic target.
		Inhibition of HIV-1 integrase by dinucleotides and trinucleotides.
		integrase in the presence of 132, 60, 30, 15, and 7.5 µ
	molpharm.aspetjournals.org /cgi/content/full/51/4/567 (4065 words)

	Establishing Proof of Concept for a New Class of Anti-HIV Drugs: The Integrase Inhibitors (Site not responding. Last check: 2007-10-08)
		Merck's L-000870810 is a novel HIV-1 integrase transfer inhibitor with potent in vitro anti-HIV activity.
		Integrase is an enzyme that HIV uses to insert its DNA into the DNA genome of human cells.
		Development of the new integrase inhibitor JT-3003 is being undertaken by Gilead Sciences, headquartered in Foster City, CA.
	www.hivandhepatitis.com /recent/experimental_drugs/032805_a.html (623 words)

	Metal-Dependent Inhibition of HIV-1 Integrase by {beta}-Diketo Acids and Resistance of the Soluble Double-Mutant ...
		Comparison of HIV-1 integrase inhibition by 5CITEP and DKA1 in the presence of Mn and Mg A, sequence of the double-stranded oligonucleotide used as substrate and schematic representation of the in vitro integration assay.
		Bischerour J, Leh H, Deprez E, Brochon JC, and Mouscadet JF (2003) Disulfide-linked integrase oligomers involving c280 residues are formed in vitro and in vivo but are not essential for human immunodeficiency virus replication.
		Young SD and the HIV integrase inhibitor discovery team (2002) L-870,810: A potent antiviral HIV integrase inhibitor with potential clinical utility XIV International AIDS Conference; 2002 Jul 7–12; Barcelona, Spain.
	molpharm.aspetjournals.org /cgi/content/full/64/3/600 (5762 words)

	DG DISPATCH - DART: Strand Transfer Specific Inhibitors Of HIV-1 Integrase May Provide Alternative HIV Treatment (Site not responding. Last check: 2007-10-08)
		Integrase processes complementary DNA and integrates it into the host cell DNA, but there are at present no inhibitors for HIV integrase.
		Using a specific assay for the inhibition of integrase, the team at Merck found a series of diketo acids (such as L-731,988) that specifically block integrase.
		Two other enzymatic functions of the integrase protein, assembly of pre-integration complexes and 3' processing of the viral DNA ends, are not inhibited by these experimental drugs.
	www.docguide.com /dg.nsf/PrintPrint/35EAF05CB5263C7C852569C3006C53B2 (452 words)

	Resolvase-like recombination performed by the TP901-1 integrase -- Breüner and Hammer 147 (8): 2051 -- Microbiology
		Overproduction of the TP901-1 integrase was induced in a culture
		Relative amounts of integrase extract added, and the positions of the bound and free fragments on the gels, are indicated.
		that decrease the affinity of the integrase for the fragment
	mic.sgmjournals.org /cgi/content/full/147/8/2051 (5290 words)

	56. A C-Terminal Nuclear Localization Signal in HIV-1 Integrase Mediates Viral Nuclear Import (Site not responding. Last check: 2007-10-08)
		Background: HIV-1 integrase, in addition to catalyzing the insertion of reverse-transcribed viral DNA into the host-cell genome, may also facilitate reverse transcription and viral DNA nuclear import.
		Fusion of a region of integrase containing the NLS to otherwise cytoplasmic proteins results in nuclear localization of the chimeras.
		Conclusions: These results indicate that the integrase NLS plays a significant role in the nuclear import of the HIV-1 genome.
	www.retroconference.org /2002/Abstract/13948.htm (222 words)

	Salk Institute for Biological Studies (Site not responding. Last check: 2007-10-08)
		Integrase is the only HIV enzyme not targeted by current drugs; reverse transcriptase and protease are blocked by drugs such as AZT and the protease inhibitors.
		The Salk team identified a pocket on the enzyme’s active surface that bore a negative charge and tested a number of small positively charged molecules for ability to bind to the pocket.
		The integrase enzyme only acts at one point in the viral life cycle — the point at which the viral genome is inserted into a cellular chromosome.
	www.salk.edu /news/releases/details.php?id=19 (538 words)

	Critical contacts between HIV-1 integrase and viral DNA identified by structure-based analysis and photo-crosslinking (Site not responding. Last check: 2007-10-08)
		Integrase is a three domain multimeric protein, that contains conserved amino acid sequence motifs in both the N-terminus (HHCC) and core domain (D,D-35-E).
		Structural analysis of the core domain (Dyda et al., 1994; Bujacz et al., 1995) revealed that integrase is structurally related to a diverse superfamily of polynucleotidyl transferases.
		M of each protein was incubated with 5 nM radiolabeled DNA for 30 min at 4°C, and then crosslinked by irradiating for 20 min at 254 nm.
	www.nature.com /emboj/journal/v16/n22/fig_tab/7590655ft.html (974 words)

	Phage TP901-1 Site-Specific Integrase Functions in Human Cells -- Stoll et al. 184 (13): 3657 -- The Journal of ...
		The TP901-1 integrase gene was PCR amplified from pCMV-TPInt
		The TP901-1 integrase was relatively sensitive to temperature
		be involved in the integrase binding and recombination function.
	jb.asm.org /cgi/content/full/184/13/3657 (4148 words)

	News Article (Site not responding. Last check: 2007-10-08)
		Striking similarities between HIV integrase and RAG1/2, a human enzyme that controls an essential step in the development of the immune system, have raised the possibility that inhibitors of HIV integrase could interfere with this vital process.
		The latest inhibitors of HIV integrase to be developed are the diketo acids, which show antiretroviral activity in cell culture and appear to be far more selective for integrase than earlier candidates.
		Although HIV integrase was more sensitive than RAG1/2 to these agents, both the compounds tested were shown to be capable of preventing the proper interaction of RAG1/2 and its target DNA sequences.
	www.mediscover.net /related.cfm?Hnid=505 (385 words)

	High-level expression of active HIV-1 integrase from a synthetic gene in human cells -- CHEREPANOV et al. 14 (10): 1389 ... (Site not responding. Last check: 2007-10-08)
		HIV-1 integrase is a protein of 32 kDa composed of three functional domains.
		Integrase was detected by Western blotting using polyclonal antibodies against HIV-1 integrase and the ECL+ system.
		the inactivating D64V mutation in the integrase gene of an HIV-1
	www.fasebj.org /cgi/content/full/14/10/1389 (6067 words)

	Mutations in the C-terminal domain of ALSV (Avian Leukemia and Sarcoma Viruses) integrase alter the concerted DNA ... (Site not responding. Last check: 2007-10-08)
		Mutations in the C-terminal domain of ALSV (Avian Leukemia and Sarcoma Viruses) integrase alter the concerted DNA integration process in vitro -- Moreau et al.
		Integrase (IN) is the retroviral enzyme responsible for the
		Chiu, R. and Grandgenett, D.P. (2003) Molecular and genetic determinants of rous sarcoma virus integrase for concerted DNA integration.
	www.ejbiochem.org /cgi/content/full/270/22/4426 (7881 words)

	C&EN: TODAY'S HEADLINES - HIV integrase-DNA binding model
		he first detailed molecular binding model consistent with experimental data on the complex of HIV integrase and viral DNA has been obtained by James M. Briggs of the University of Houston and coworkers [J.
		HIV integrase binds to a DNA copy of the virus's genome and catalyzes its insertion into a host chromosome, inducing the host cell to synthesize viral proteins.
		The enzyme is thus an essential part of the viral life cycle and a target for the design of therapeutic inhibitors.
	pubs.acs.org /cen/today/8204binding.html (172 words)

	Construction of Transgenic Drosophila by Using the Site-Specific Integrase From Phage {phi}C31 -- Groth et al. 166 (4): ...
		of the integrase plasmid and/or 400 ng of the donor plasmid.
		The integrase was induced at 24 hr by the
		integrase in bacteria and that the recombination occurred in
	www.genetics.org /cgi/content/full/166/4/1775 (3591 words)

	Mutations Resist New HIV Integrase Inhibitor (Site not responding. Last check: 2007-10-08)
		As the result of the selection process, nine mutations arose in the catalytic domain of integrase.
		However, the researchers note that all mutants remained susceptible to the integrase DNA binding inhibitor V-165.
		Given this, the researchers conclude that use of integrase strand transfer inhibitors along with integrase DNA binding inhibitors and current antiretroviral compounds, "is of potential interest when designing the next generation of anti-HIV drugs."
	www.hivandhepatitis.com /recent/resistance/112204_f.html (184 words)

	High-level expression of active HIV-1 integrase from a synthetic gene in human cells -- CHEREPANOV et al. 14 (10): 1389 ...
		High-level expression of active HIV-1 integrase from a synthetic gene in human cells -- CHEREPANOV et al.
		High-level expression of active HIV-1 integrase from a synthetic gene in human cells
		integrase from a synthetic gene in human cells.
	www.fasebj.org /cgi/content/abstract/14/10/1389 (700 words)

	Molecular Dynamics Studies on the HIV-1 Integrase Catalytic Domain -- Lins et al. 76 (6): 2999 -- Biophysical Journal
		The catalytic domain of avian sarcoma virus integrase: conformation of the active-site residues in the presence of divalent cations.
		The core and carboxyl-terminal domains of the integrase protein of human immunodeficiency virus type 1 each contribute to non-specific DNA binding.
		Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/retrotransposon integrases and bacterial insertion sequence transposons.
	www.biophysj.org /cgi/content/full/76/6/2999 (5864 words)

	HIV-1 Integrase (Site not responding. Last check: 2007-10-08)
		The human immunodeficiency virus (HIV-1) uses an enzyme, a so-called integrase, to carry out the integration of its viral DNA into the host chromosome thereby tricking the host cell machinery into making viral proteins.
		The HIV integrase is composed of three domains, as shown below.
		It is thought that the HIV and ASV integrases might operate via a two metal mechanism like that proposed by Beese and Steitz for DNA polymerase I (Beese L.S., Steitz T.A. Structural basis for the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I: a two metal ion mechanism.
	adrik.bchs.uh.edu /integrase.html (232 words)

	Fox Chase Cancer Center: Anna Marie Skalka, Ph.D. (Site not responding. Last check: 2007-10-08)
		To address such questions, we study the integrase proteins of the avian sarcoma and leukosis virus (ASLV) and the human immunodeficiency virus type 1 (HIV-1).
		Mechanistic aspects of ASV integrase catalytic activity and structural analyses of the integrase proteins of both viruses are being performed in collaboration with Isaac Wong, at Oregon State University, and with the crystallography laboratory of Alex Wlodawer at NCI.
		A second, equally active area of research in our laboratory centers on integration-related events that take place inside of infected cells where integrase functions as a component of a large nucleo- protein complex that is formed in the cytoplasm shortly after infection.
	www.fccc.edu /research/pid/skalka (908 words)

	ipedia.com: Retrovirus Article (Site not responding. Last check: 2007-10-08)
		It relies on reverse transcriptase to perform a kind of reverse transcription of its genome from RNA into DNA for insertion by integrase i...
		It relies on reverse transcriptase to perform a kind of reverse transcription of its genome from RNA into DNA for insertion by integrase into the host's genome.
		The virus itself is just a storage form for its RNA; the reverse transcription takes place in the host's cytosol.
	www.ipedia.com /retrovirus.html (230 words)

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