Integrase and integration: biochemical activities of HIV-1 integrase

Integration of retroviral DNA is an obligatory step of retrovirus replication because proviral DNA is the template for productive infection. Integrase, a retroviral enzyme, catalyses integration. The process of integration can be divided into two sequential reactions. The first one, named 3'-processing, corresponds to a specific endonucleolytic reaction which prepares the viral DNA extremities to be competent for the subsequent covalent insertion, named strand transfer, into the host cell genome by a trans-esterification reaction. Recently, a novel specific activity of the full length integrase was reported, in vitro, by our group for two retroviral integrases (HIV-1 and PFV-1). This activity of internal cleavage occurs at a specific palindromic sequence mimicking the LTR-LTR junction described into the 2-LTR circles which are peculiar viral DNA forms found during viral infection. Moreover, recent studies demonstrated the existence of a weak palindromic consensus found at the integration sites. Taken together, these data underline the propensity of retroviral integrases for binding symmetrical sequences and give perspectives for targeting specific sequences used for gene therapy

Randomized field trial on the effects of body weight and short transport on stress and immune variables in 2‐ to 4‐week‐old dairy calves

BACKGROUND: Whether underweight calves respond differently to transport stress, enhancing their disease risk, is currently unknown. OBJECTIVE: To determine the effects of low body weight and transport stress on immune variables. ANIMALS: Twenty-one 2- to 4-week-old male Holstein calves, housed on a commercial farm. METHODS: Randomized clinical trial. Full factorial design with 4 treatment groups: low body weight (≤46 kg)/no transport (LOWCON); low body weight/transport (LOWTRANS); normal body weight (>46 kg)/no transport (NORMCON), and normal body weight/transport (NORMTRANS). Transport duration was 2 hours. RESULTS: Transport significantly increased serum cortisol concentration (77.8 μg/mL; 95% confidence interval [CI], 37.8-131.6; P < .001), interleukin (IL)-17A (344.9 pg/mL; 95% CI, 32.2-556.5; P = .04), and tumor necrosis factor-α (TNF-α) (218.2 pg/mL; 95% CI, 32.5-368.3; P = .03) production after lipopolysaccharide (LPS) stimulation. Body weight did not affect any of the studied variables. However, the interaction of transport and body weight was significant. LOWTRANS calves showed increased monocyte count (2.0 × 109 /L; 95% CI, 0.6-4.2; P < .05) and interleukin IL-17A production (106.0 pg/mL; 95% CI, 4.2-306.9; P = .03) compared to normal weight calves and increased TNF-α production (275.6 pg/mL; 95% CI, 2.6-463.0; P = .02) compared to LOWCON calves in unstimulated peripheral blood mononuclear cells (PBMCs) after transport. CONCLUSION AND CLINICAL IMPORTANCE: These findings contribute to our understanding of increased disease susceptibility of underweight calves when transported. Gamma globulin concentration was identified as important interfering factor in studies on immune variables in neonatal calves

Effects of short transport and body condition on stress and immune parameters in 2 to-4-week old dairy calves

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Efficient and Specific Internal Cleavage of a Retroviral Palindromic DNA Sequence by Tetrameric HIV-1 Integrase

BACKGROUND: HIV-1 integrase (IN) catalyses the retroviral integration process, removing two nucleotides from each long terminal repeat and inserting the processed viral DNA into the target DNA. It is widely assumed that the strand transfer step has no sequence specificity. However, recently, it has been reported by several groups that integration sites display a preference for palindromic sequences, suggesting that a symmetry in the target DNA may stabilise the tetrameric organisation of IN in the synaptic complex. METHODOLOGY/PRINCIPAL FINDINGS: We assessed the ability of several palindrome-containing sequences to organise tetrameric IN and investigated the ability of IN to catalyse DNA cleavage at internal positions. Only one palindromic sequence was successfully cleaved by IN. Interestingly, this symmetrical sequence corresponded to the 2-LTR junction of retroviral DNA circles-a palindrome similar but not identical to the consensus sequence found at integration sites. This reaction depended strictly on the cognate retroviral sequence of IN and required a full-length wild-type IN. Furthermore, the oligomeric state of IN responsible for this cleavage differed from that involved in the 3'-processing reaction. Palindromic cleavage strictly required the tetrameric form, whereas 3'-processing was efficiently catalysed by a dimer. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that the restriction-like cleavage of palindromic sequences may be a general physiological activity of retroviral INs and that IN tetramerisation is strongly favoured by DNA symmetry, either at the target site for the concerted integration or when the DNA contains the 2-LTR junction in the case of the palindromic internal cleavage