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MukB colocalizes with the oriC region and is required for organization of the two Escherichia coli chromosome arms into separate cell halves 总被引:1,自引:1,他引:0
Danilova O Reyes-Lamothe R Pinskaya M Sherratt D Possoz C 《Molecular microbiology》2007,65(6):1485-1492
The circular Escherichia coli chromosome is organized by bidirectional replication into two equal left and right arms (replichores). Each arm occupies a separate cell half, with the origin of replication (oriC) at mid-cell. E. coli MukBEF belongs to the ubiquitous family of SMC protein complexes that play key roles in chromosome organization and processing. In mukBEF mutants, viability is restricted to low temperature with production of anucleate cells, reflecting chromosome segregation defects. We show that in mukB mutant cells, the two chromosome arms do not separate into distinct cell halves, but extend from pole to pole with the oriC region located at the old pole. Mutations in topA, encoding topoisomerase I, do not suppress the aberrant positioning of chromosomal loci in mukB cells, despite suppressing the temperature-sensitivity and production of anucleate cells. Furthermore, we show that MukB and the oriC region generally colocalize throughout the cell cycle, even when oriC localization is aberrant. We propose that MukBEF initiates the normal bidirectional organization of the chromosome from the oriC region. 相似文献
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Disruption of HIV-1 integrase-DNA complexes by short 6-oxocytosine-containing oligonucleotides 总被引:1,自引:0,他引:1
Brodin P Pinskaya M Buckle M Parsch U Romanova E Engels J Gottikh M Mouscadet JF 《Biochemistry》2002,41(5):1529-1538
We recently found that oligonucleotides containing the 6-oxocytosine heterocyclic base are efficient inhibitors of the HIV-1 integrase in vitro [Brodin, P., et al. (2001) Nucleosides Nucleotides Nucleic Acids 20, 481-486]. In this report, we demonstrate that the inhibition arises from a noncompetitive mechanism in which the modified oligonucleotide attacks the integrase-DNA complex, leading to its active disruption. This conclusion is based on the following results. First, despite the fact that the respective affinities of a 6-oxocytosine-containing oligonucleotide and of its nonmodified counterpart for integrase were identical, only the modified compound inhibited the enzyme activities. Second, DNA binding and UV cross-linking assays indicated that the 6-oxocytosine-containing oligonucleotide prevented the formation of a stable integrase-DNA complex. Third, the kinetics of the dissociation of the integrase-DNA complex were dramatically accelerated in the presence of the modified ODN, whereas the nonmodified counterpart did not influence the dissociation. This mechanism was supported by the ability of the 6-oxocytosine-containing oligonucleotide to inhibit the strand transfer activity of HIV-1 preintegration complexes in vitro. Disruption of integrase-DNA complexes by 6-oxocytosine-containing oligonucleotides constitutes an original mechanism of integration inhibition, therefore suggesting a strategy for searching for inhibitors of the HIV-1 preintegration complexes. 相似文献
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HIV-1 integrase complexes with DNA dissociate in the presence of short oligonucleotides conjugated to acridine 总被引:2,自引:0,他引:2
Pinskaya M Romanova E Volkov E Deprez E Leh H Brochon JC Mouscadet JF Gottikh M 《Biochemistry》2004,43(27):8735-8743
The human immunodeficiency virus type 1 (HIV-1) integrase is an essential enzyme in the life cycle of the virus and is therefore an attractive target for the development of new antiviral drugs. Among them, inhibitors which are capable of targeting the preassembled integrase/DNA complex are of particular interest, because they could suppress integrase activity in the context of the HIV-1 preintegration complex. Here, we study the mechanism of action of 11-mer oligonucleotides, which are efficient inhibitors of the catalytic activity of integrase, provided that they are conjugated to a hydrophobic compound, acridine. To understand the mechanism of the conjugate inhibitory action, we used a steady-state fluorescence anisotropy assay, which allowed us to study the stability of the integrase/DNA complex in various conditions. We found that oligonucleotide-acridine conjugates induced the efficient dissociation of preassembled integrase/DNA complexes. The simultaneous presence of both acridine and an oligonucleotidic moiety is required for the inhibitory activity of conjugates. However, the dissociation effect is not dependent on the oligonucleotide sequence. Finally, our results suggest that the conjugates bind directly to integrase within its complex with DNA at a site different from the viral DNA binding site. 相似文献
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Brodin P Pinskaya M Parsch U Bischerour J Leh H Romanova E Engels JW Gottikh M Mouscadet JF 《Nucleosides, nucleotides & nucleic acids》2001,20(4-7):481-486
Integration of the proviral DNA into the genome of infected cells is a key step of HIV-1 replication. Integration is catalyzed by the viral enzyme integrase (IN). 6-oxocytidine-containing oligonucleotides were found to be efficient inhibitors of integrase in vitro. The inhibitory effect is sequence-specific and strictly requires the presence of the 6-oxocytidine base. It is due to the impairment of the integrase binding to its substrate and does not involve an auto-structure of the oligonucleotide. 相似文献
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Branched oligonucleotide-intercalator conjugate forming a parallel stranded structure inhibits HIV-1 integrase. 总被引:2,自引:0,他引:2
P Brodin M Pinskaya E Volkov E Romanova H Leh C Auclair J F Mouscadet M Gottikh 《FEBS letters》1999,460(2):270-274
Integration of a DNA copy of the HIV-1 genome into chromosomal DNA of infected cells is a key step of viral replication. Integration is carried out by integrase, a viral protein which binds to both ends of viral DNA and catalyses reactions of the 3'-end processing and strand transfer. A 3'-3' branched oligonucleotide functionalised by the intercalator oxazolopyridocarbazole at each 5'-end was found to inhibit integration in vitro. We show that both a specific (G,A) sequence and the OPC intercalating agent contribute to the capability of the branched oligonucleotide to form a parallel stranded structure responsible for the inhibition. 相似文献
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M. B. Gottikh Y. I. Alekseev A. V. Perminov M. D. Pinskaya Z. A. Shabarova 《Nucleosides, nucleotides & nucleic acids》2013,32(6-7):1625-1627
Abstract The ability of oligodeoxyxylopyrimidilates to form triplexes with complementary single-stranded DNA at the neutral pH was found. The complex composition, relative strand orientation and base-pairing scheme were determined using electromobility shift assay and thermal denaturation experiments. 相似文献
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Pinskaya M. D. Brodin P. Romanova E. A. Volkov E. M. Mouscadet J.-F. Gottikh M. B. 《Molecular Biology》2000,34(6):888-894
Oligonucleotide inhibitors of the HIV-1 DNA integration identified to date are reviewed. Two basic strategies of blocking the integration are considered: shielding the integrase-binding sites on the viral DNA by triplex-forming oligonucleotides, and directly inhibiting the enzyme with oligonucleotide agents. 相似文献