The RTCs of viruses carrying the subtype B RT polymerase domain,

The RTCs of viruses carrying the subtype B RT polymerase domain, harvested at 1 h post infection displayed a 2. 5 and 5 fold Verdinexor (KPT-335)? higher relative amount of strong stop cDNA with respect to those carrying the 1084i RT polymerase domain. The ratios of early cDNA between these strains, measured at 5 h after infection, were about 2x for NL backbone and 2. 5x for 1084i backbone viruses. Similar results were observed in accu mulation of the positive strand DNA measured at 5 h post infection, suggesting that the difference in cDNA accumulation between the viruses with RTs from B and C subtypes are dependent on the initial steps of the reverse transcription.

Taken together our data indicate that the presence of the RT, as well as only the polymerase, or the connec tion and RNase H domains of RT from subtype C viruses leads to a lower level of accumulation of strong stop cDNA and late reverse transcription products, in both intact virions and intracytoplasmic Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries RTCs indepen dent of the virus backbone. The difference in viral DNA accumulation between viruses carrying RT from subtype B and C isolates may eventually determine the overall level of viral replication, that is consistent with the pub lished data on subtype associated effect of RT on viral replicative fitness. Cells infected with viruses Inhibitors,Modulators,Libraries carrying RT functional domains from HIV 1 subtype C isolates display decreased viral DNA integration Lower levels of accumulation of reverse transcription products in viruses carrying subtype C pol products may correlate with the level of viral DNA integration into the host chromosomes.

We then analyzed integration of these viruses using a two step Alu based nested PCR assay. Quantitative analysis of the cellular DNA showed that viruses carrying protease and RT polymer ase domains from different subtype C isolates, NLpolL, NLpolL and NLpolL, displayed between three to fifty fold fewer proviruses than sub type B NL4 3. To further confirm that this Inhibitors,Modulators,Libraries difference is due to the functional domains of RT, we compared various recombinant viruses that carry only the polymerase domain from subtype B or subtype C isolates with virus strains carrying the whole Pol fragment without protease, or the connection, RNase H, and the integrase sequences from Inhibitors,Modulators,Libraries subtype B and C iso lates. As expected, subtype B NL RTpd had simi lar levels of integrated provirus as NL4 3.

The two viruses carrying subtype C RT polymerase inhibitor Rapamycin domain had 2 2. 5 fold lower levels of integration at 24 h and 3 and 4 fold lower at 48 h post infection. These findings are consistent with our data on cDNA accumulation in the virions and RTCs. Our results also showed that the integrase from B and C subtypes did not significantly affect the integration rate of the viruses containing B and C RT domains. Ana lysis performed at 48 h post infection showed a mean of threefold higher levels of integration than at 24 h post infection.

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