In vivo evidence that TATA-binding protein/SL1 colocalizes with UBF and RNA polymerase I when rRNA synthesis is either active or inactive.
Here we show that the TATA-binding protein (TBP) is localized in the nucleoplasm and in the nucleolus of mammalian cells, consistent with its known involvement in transcription by RNA polymerase I, II, and III. In the nucleolus of actively growing cells, TBP colocalizes with upstream binding factor (UBF) and RNA polymerase I at the sites of rRNA transcription. During mitosis, when rRNA synthesis is down-regulated, TBP colocalizes with TBP-associated factors for RNA polymerase I (TAF(I)s), UBF, and RNA polymerase I on the chromosomal regions containing the rRNA genes. Treatment of cells with a low concentration of actinomycin D inhibits rRNA synthesis and causes a redistribution of the rRNA genes that become concentrated in clusters at the periphery of the nucleolus. A similar redistribution was observed for the major components of the rRNA transcription machinery (i.e., TBP, TAF(I)s, UBF, and RNA polymerase I), which still colocalized with each other. Furthermore, anti-TBP antibodies are shown to coimmunoprecipitate TBP and TAF(I)63 in extracts prepared from untreated and actinomycin D-treated cells. Collectively, the data indicate that in vivo TBP/promoter selectivity factor, UBF, and RNA polymerase I remain associated with both active and inactive rRNA genes.
Jordan
P
P
Institute of Histology and Embryology, Faculty of Medicine, University of Lisbon, Portugal.
Mannervik
M
M
Tora
L
L
Carmo-Fonseca
M
M
D016475:3T3 Cells; D000818:Animals; D002466:Cell Nucleolus / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000032:analysis*; D003609:Dactinomycin / Q000494:pharmacology; D005796:Genes; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008938:Mitosis; D035463:Pol1 Transcription Initiation Complex Proteins; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012318:RNA Polymerase I / Q000032:analysis*; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D035181:TATA-Box Binding Protein; D014157:Transcription Factors / Q000032:analysis*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D035463:Pol1 Transcription Initiation Complex Proteins; D011500:Protein Synthesis Inhibitors; D012335:RNA, Ribosomal; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; C063844:transcription factor UBF; C063660:transcription initiation factor TIF-IB; D003609:Dactinomycin; D012318:RNA Polymerase I