Vels of viral particles, we speculate that average levels of host-viral chimeric transcripts in these cells exceed those in the two J-Lat cell lines. These results thus suggest that TI plays a key role in primary CD4+ T cells. In J-Lat 9.2 and 15.4 cells, the host-viral chimeric transcripts terminated at the pA site in the 5'LTR, but transcripts initiating at the host promoter and terminating at t
T the level of gene expression act cooperatively to establish and maintain HIV latency. In conclusion, our study reveals that TI represents a key mechanism that antagonizes proviral gene expression to promote the latency of HIV. Furthermore, it demonstrates several means that could be used to counteract TI for reactivating latent HIV. Future mechanistic studies linking TI and other transcriptional
Ipt Author Manuscript Author Manuscript Author ManuscriptExtended Data Figure 2. Development of anti-MAb immune responses in recipients of antiHIV-1 bNAbsNature. Author manuscript; available in PMC 2016 November 29.Gautam et al.Pagea, b, c, d, Longitudinal analysis of anti-VRC01, anti-3BNC117, anti-10-1074 and antiVRC01-LS antibody responses respectively, following a single intravenous infusion of
One (Figure 7D, bars 2 and 6) and in combination with HMBA (Figure 7D, bars 3 and 7) or Tat (Figure 7D, bars 4 and 8) to levels in untreated cells (Figure 7D, bars 1 and 5). Interestingly, *mRNA levels decreased concomitantly with the increased activation of transcription from the 5'LTR in both cell lines. Probably, the formation of the PIC at the 5'LTR prevents elongating complex from the upstrea
One (Figure 7D, bars 2 and 6) and in combination with HMBA (Figure 7D, bars 3 and 7) or Tat (Figure 7D, bars 4 and 8) to levels in untreated cells (Figure 7D, bars 1 and 5). Interestingly, *mRNA levels decreased concomitantly with the increased activation of transcription from the 5'LTR in both cell lines. Probably, the formation of the PIC at the 5'LTR prevents elongating complex from the upstrea
Ed cells. Furthermore, TNF- stimulation led to the appearance of Sp1 and initiating RNAPII on the 5'LTR and additional enrichments of initiating and elongating RNAPII on the 3'LTR. These effects are most likely achieved through the activation of NF-B, which stimulates several steps of viral transcription including the PIC formation, transcription initiation and transcription elongation. Thus, our
Ranscription units (Han et al., 2004; Lewinski et al., 2005). In the present work, we extended this knowledge by a detailed analysis of how TI impacts viral transcription in latent cells. Although TI occurs regardless of the orientation of the two promoters, we studied TI phenomenon in two J-Lat cell lines with the same orientation of the host and viral promoters, which allowed us to detect and an
Ly 20 activation of viral transcription (see Figure 7) already precludes detection of the upstream transcription. This finding is not surprising, as we demonstrated that there is 11-fold or 38-fold more viral mRNA than *mRNA in TNF- treated cells (Figure 2D, bars 3 and 4). Predictably, our control experiments demonstrate that the ratio between the Rev- and Env-containing transcripts was 1 in untr