The species-specific RNA polymerase I transcription factor SL-1 binds to upstream binding factor.
Transcription of the 45S rRNA genes is carried out by RNA polymerase I and at least two trans-acting factors, upstream binding factor (UBF) and SL-1. We have examined the hypothesis that SL-1 and UBF interact. Coimmunoprecipitation studies using an antibody to UBF demonstrated that TATA-binding protein, a subunit of SL-1, associates with UBF in the absence of DNA. Inclusion of the detergents sodium dodecyl sulfate and deoxycholate disrupted this interaction. In addition, partially purified UBF from rat cell nuclear extracts and partially purified SL-1 from human cells coimmunoprecipitated with the anti-UBF antibody after mixing, indicating that the UBF-SL-1 complex can re-form. Treatment of UBF-depleted extracts with the anti-UBF antibody depleted the extracts of SL-1 activity only if UBF was added to the extract prior to the immunodepletion reaction. Furthermore, SL-1 activity could be recovered in the immunoprecipitate. Interestingly, these immunoprecipitates did not contain RNA polymerase I, as a monospecific antibody to the 194-kDa subunit of RNA polymerase I failed to detect that subunit in the immunoprecipitates. Treatment of N1S1 cell extracts with the anti-UBF antibody depleted the extracts of SL-1 activity but not TFIIIB activity, suggesting that the binding of UBF to SL-1 is specific and not solely mediated by an interaction between UBF and TATA-binding protein, which is also a component of TFIIIB. These data provide evidence that UBF and SL-1 interact.
Hempel
W M
WM
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822, USA.
Cavanaugh
A H
AH
Hannan
R D
RD
Taylor
L
L
Rothblum
L I
LI
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D035463:Pol1 Transcription Initiation Complex Proteins; D011233:Precipitin Tests; D011485:Protein Binding; D012318:RNA Polymerase I / Q000378:metabolism; D051381:Rats; D013045:Species Specificity; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D035463:Pol1 Transcription Initiation Complex Proteins; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; C063844:transcription factor UBF; C063660:transcription initiation factor TIF-IB; D012318:RNA Polymerase I