SATB1-binding sequences and Alu-like motifs define a unique chromatin context in the vicinity of human immunodeficiency virus type 1 integration sites.
ABSTRACT Retroviral integration has recently been shown to be nonrandom, favoring transcriptionally active regions of chromatin. However, the mechanism for integration site selection by retroviruses is not clear. We show here the occurrence of Alu-like motifs in the sequences flanking the reported viral integration sites that are significantly different from those obtained from the randomly picked sequences from the human genome, suggesting that unique primary sequence features exist in the genomic regions targeted by human immunodeficiency virus type 1 (HIV-1). Additionally, these sequences were preferentially bound by SATB1, the T lineage-restricted chromatin organizer, in vitro and in vivo. Alu repeats make up nearly 10% of the human genome and have been implicated in the regulation of transcription. To specifically isolate sequences flanking the viral integration sites and also harboring both Alu-like repeats and SATB1-binding sites, we combined chromatin immunoprecipitation with sequential PCRs. The cloned sequences flanking HIV-1 integration sites were specifically immunoprecipitated and amplified from the pool of anti-SATB1-immunoprecipitated genomic DNA fragments isolated from HIV-1 NL4.3-infected Jurkat T-cell chromatin. Moreover, many of these sequences were preferentially partitioned in the DNA associated tightly with the nuclear matrix and not in the chromatin loops. Strikingly, many of these regions were disfavored for integration when SATB1 was silenced, providing unequivocal evidence for its role in HIV-1 integration site selection. We propose that definitive sequence features such as the Alu-like motifs and SATB1-binding sites provide a unique chromatin context in vivo which is preferentially targeted by the HIV-1 integration machinery.
Article: Response to psychological test in elderly patients with hand-arm vibration syndrome and healthy controls.[show abstract] [hide abstract]
ABSTRACT: We investigated the parasympathetic nervous response to psychological test using heart rate variation (HRV) during deep breathing in elderly patients with hand-arm vibration syndrome and healthy controls. Average age (SD) of 16 patients with vibration-induced white finger (VWF), 13 patients without VWF and 12 healthy controls was 60.1 (2.8), 60.6 (4.2) and 58.8 (3.8), respectively. After an initial supine rest for 40 min, psychological test (Stroop color word test and mirror drawing test) was performed for 20 min. The indexes of HRV (Mean R-R, SD, RMSSD and CV) were calculated. Blood pressure and heart rate were also measured. The indexes of HRV did not differ between the groups before exposure. The SD, RMSSD and CV of the patients without VWF during supine deep breathing after 3 min post-exposure supine rest were significantly lower than those of the control group (p < 0.05). The Mean R-R of the patients without VWF significantly increased (p < 0.05). Blood pressure did not differ in either before or after exposure measurements. The results suggest that the post-exposure response of the parasympathetic nervous system to psychological test reduced in the patients without VWF.Industrial Health 10/1997; 35(4):533-6. · 0.94 Impact Factor
Article: Assessment of ammonia toxicity in tests with the microalga, Nephroselmis pyriformis, Chlorophyta.[show abstract] [hide abstract]
ABSTRACT: Previous studies of an industrial effluent have indicated toxic effects by the ammonium/water system in tests with the unicellular green alga, Nephroselmis pyriformis. This investigation was undertaken to determine the toxicity of ammonia to this alga, and to identify the dominant toxicant in an industrial process effluent. The algal standard test was modified to improve the pH control, i.e. by testing at shorter exposure time and in the presence of a buffering agent, 3 mM HEPES. Ammonia was found to be the predominant toxic form in the dissociating ammonium/water system and the specific toxicity of ammonia (EC50, 24 h exposure) was 2.34 microM (32.8 microg ammonia nitrogen/L). Due to the dissociation, the toxicity is strongly pH dependent. Joint toxicity with additive effects of ammonia and ammonium ions was indicated, but the toxicity of ammonium ions was almost a factor 100 less (EC50, 24 h exposure, 224 microM or 3140 microg ammonium-nitrogen/L). According to the results the EC50 for effect on growth rate of N. pyriformis in seawater of pH 8.0 is 71 microM total ammonium. Hence, this alga appears to be more sensitive to ammonia than other studied marine plankton algae. Ammonia was found to be the dominant toxicant in the industrial effluent, using N. pyriformis as test organism.Water Research 03/2003; 37(3):477-84. · 4.86 Impact Factor
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ABSTRACT: A central feature of retrovirus replication is integration of the provirus into host cell DNA, but the specificity of this step for cell target sequences has not been clarified. To investigate this issue, we developed a method for screening and comparing large numbers of unselected integration events. Using a replication-competent Rous sarcoma virus containing a bacterial suppressor tRNA gene as a selectable marker, we obtained collections of clones comprising integrated provirus together with host flanking sequences. Hybridization and sequence analysis of the flanking sequence reveals the presence of a number of strongly preferred integration targets. Within these targets, independent integration events occur at sites identical to the base.Cell 06/1988; 53(4):531-7. · 32.40 Impact Factor