The transcript release factor PTRF augments ribosomal gene transcription by facilitating reinitiation of RNA polymerase I.
Termination of murine rDNA transcription by RNA polymerase I (Pol I) requires pausing of Pol I by terminator-bound TTF-I (transcription termination factor for Pol I), followed by dissociation of the ternary complex by PTRF (Pol I and transcript release factor). To examine the functional correlation between transcription termination and initiation, we have compared transcription on terminator-containing and terminator-less rDNA templates. We demonstrate that terminated RNA molecules are more efficiently synthesized than run-off transcripts, indicating that termination facilitates reinitiation. Transcriptional enhancement is observed in multiple- but not single-round transcription assays measuring either promoter-dependent or promoter-independent Pol I transcription. Increased synthesis of terminated transcripts is observed in crude extracts but not in a PTRF-free reconstituted transcription system, indicating that PTRF-mediated release of pre-rRNA is responsible for transcriptional enhancement. Consistent with PTRF serving an important role in modulating the efficiency of rRNA synthesis, PTRF exhibits pronounced charge heterogeneity, is phosphorylated at multiple sites and fractionates into transcriptionally active and inactive forms. The results suggest that regulation of PTRF activity may be an as yet unrecognized means to control the efficiency of ribosomal RNA synthesis.
Jansa
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Division of Molecular Biology of the Cell II, German Cancer Research Center, D-69120 Heidelberg, Germany.
Burek
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C
Sander
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Grummt
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I
D016475:3T3 Cells; D000818:Animals; D004275:DNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins; D051379:Mice; D010766:Phosphorylation; D012318:RNA Polymerase I / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D012270:Ribosomes / Q000201:enzymology / Q000235:genetics*; D013698:Templates, Genetic; D014157:Transcription Factors; D014158:Transcription, Genetic
C112745:Cavin1 protein, mouse; D004275:DNA, Ribosomal; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D012322:RNA Precursors; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D014157:Transcription Factors; C504004:Ttf1 protein, mouse; D012318:RNA Polymerase I