7SK. For example, the single-stranded RNA binding MGCD-0103 site proteins 3’UTR Activates Transcription r luciferase activity. Data are represented as firefly/Renilla activity normalized to the sample without Tat. Cell extracts were immunoblotted and probed with antibody to CDK9. HIC 3’UTR specifically releases 7SK RNA from P-TEFb. Immunoprecipitated P-TEFb was assayed for 7SK RNA as in panel 14985929 B. The number of PCR cycles is specified. doi:10.1371/journal.pone.0001010.g005 hnRNPA1 and A2 bind exclusively to stem 3 of 7SK while hnRNP R and Q1 bind stems 1 and 3. Stem 1 is also important for the binding of RHA. An extensive structure-function study led to the demonstration that both the 59- and 39-terminal stem-loop structures of 7SK are required for binding P-TEFb in vivo. The 59 stem-loop structure is necessary for binding HEXIM1, which is a prerequisite for binding P-TEFb; in the next step, the 39-terminal hairpin of 7SK interacts with P-TEFb. We therefore used bioinformatic tools to examine the terminal segment of 
the HIC 3’UTR for the existence of an imperfect stem-loop structure that resembles stem 4 of 7SK. A similar hairpin structure is predicted within the HIC 3’UTR. The loop region of this hairpin contains a sequence, AUPuUGG, that is shared with stem 4 of 7SK RNA. A neighboring sequence in the HIC 3’UTR is predicted to form 6 3’UTR Activates Transcription P-TEFb. Thus, P-TEFbcontaining RNPs could have a role in various aspects of RNA processing as well as in transcription itself. HIC protein and mRNA We initially isolated HIC as a protein that 14985929 binds the cyclin T1 component of P-TEFb. Subsequently, we found that overexpression of its I-mfa domain inhibits P-TEFb-dependent gene expression. In addition, HIC possesses RNA binding activity via its conserved basic region . It is tempting to speculate that the HIC protein and the 3’UTR of its mRNA interact with P-TEFb in a fashion reminiscent of the Tat/TAR/P-TEFb and HEXIM/7SK/PTEFb interactions. In these well-studied instances, RNA molecules are key participants in the positive and negative regulation of PTEFb function, respectively. Interestingly Tat was recently demonstrated to bind 7SK and disrupt the inhibitory P-TEFb complex. It remains to be determined whether HIC can also bind to 7SK and disrupt its interaction with HEXIM, thereby releasing P-TEFb. While the physiological roles of HIC are not yet well understood, its expression, like that of its Xenopus ortholog XIC, appears to be under tight control . The HIC and XIC proteins, which are active during differentiation and developmental scenarios, could activate P-TEFb via interactions with cyclin T1 and the HIC 3’UTR causing a reduction of the inhibitory complex. Conceivably this action could be restricted to the vicinity of the transcription apparatus at specific genes. Documentation of the existence of such complexes is an important next step in evaluating this model. Finally, our findings suggest a general mechanism whereby gene expression can be controlled at the level of P-TEFb by signals transmitted through mRNA as well as protein modulators. Exploitation of this phenomenon to increase transcription and gene expression by transfection of the HIC 3’UTR, or its derivatives, could be useful when enhanced yields of RNA or protein is required either in commercial or laboratory settings. a structure similar to that predicted for stem 2 of 7SK which contributes to, but is not essential for, its binding to P-TEFb. Both structures are present in
