Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription

Sandy Hung, Analia Lesmana, Abigail Peck, Rachel Lee, Elly Tchoubrieva, Katherine M Hannan, Jane I Lin, Karen E Sheppard, Katarzyna Jastrzebski, Leonie M Quinn, Lawrence I Rothblum, Richard B. Pearson, Ross D. Hannan, Elaine Sanij

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Transcription of the ribosomal RNA genes (rDNA) by RNA polymerase I (Pol I) is a major control step for ribosome synthesis and is tightly linked to cellular growth. However, the question of whether this process is modulated primarily at the level of transcription initiation or elongation is controversial. Studies in markedly different cell types have identified either initiation or elongation as the major control point. In this study, we have re-examined this question in NIH3T3 fibroblasts using a combination of metabolic labeling of the 47S rRNA, chromatin immunoprecipitation analysis of Pol I and overexpression of the transcription initiation factor Rrn3. Acute manipulation of growth factor levels altered rRNA synthesis rates over 8-fold without changing Pol I loading onto the rDNA. In fact, robust changes in Pol I loading were only observed under conditions where inhibition of rDNA transcription was associated with chronic serum starvation or cell cycle arrest. Overexpression of the transcription initiation factor Rrn3 increased loading of Pol I on the rDNA but failed to enhance rRNA synthesis in either serum starved, serum treated or G0/G1 arrested cells. Together these data suggest that transcription elongation is rate limiting for rRNA synthesis. We propose that transcription initiation is required for rDNA transcription in response to cell cycle cues, whereas elongation controls the dynamic range of rRNA synthesis output in response to acute growth factor modulation.

Original languageEnglish
Pages (from-to)36-48
Number of pages13
JournalGene
Volume612
DOIs
Publication statusPublished - 15 May 2017
Externally publishedYes

Keywords

  • rDNA
  • RNA polymerase I
  • Rrn3

Cite this

Hung, S., Lesmana, A., Peck, A., Lee, R., Tchoubrieva, E., Hannan, K. M., ... Sanij, E. (2017). Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription. Gene, 612, 36-48. https://doi.org/10.1016/j.gene.2016.12.015
Hung, Sandy ; Lesmana, Analia ; Peck, Abigail ; Lee, Rachel ; Tchoubrieva, Elly ; Hannan, Katherine M ; Lin, Jane I ; Sheppard, Karen E ; Jastrzebski, Katarzyna ; Quinn, Leonie M ; Rothblum, Lawrence I ; Pearson, Richard B. ; Hannan, Ross D. ; Sanij, Elaine. / Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription. In: Gene. 2017 ; Vol. 612. pp. 36-48.
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abstract = "Transcription of the ribosomal RNA genes (rDNA) by RNA polymerase I (Pol I) is a major control step for ribosome synthesis and is tightly linked to cellular growth. However, the question of whether this process is modulated primarily at the level of transcription initiation or elongation is controversial. Studies in markedly different cell types have identified either initiation or elongation as the major control point. In this study, we have re-examined this question in NIH3T3 fibroblasts using a combination of metabolic labeling of the 47S rRNA, chromatin immunoprecipitation analysis of Pol I and overexpression of the transcription initiation factor Rrn3. Acute manipulation of growth factor levels altered rRNA synthesis rates over 8-fold without changing Pol I loading onto the rDNA. In fact, robust changes in Pol I loading were only observed under conditions where inhibition of rDNA transcription was associated with chronic serum starvation or cell cycle arrest. Overexpression of the transcription initiation factor Rrn3 increased loading of Pol I on the rDNA but failed to enhance rRNA synthesis in either serum starved, serum treated or G0/G1 arrested cells. Together these data suggest that transcription elongation is rate limiting for rRNA synthesis. We propose that transcription initiation is required for rDNA transcription in response to cell cycle cues, whereas elongation controls the dynamic range of rRNA synthesis output in response to acute growth factor modulation.",
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Hung, S, Lesmana, A, Peck, A, Lee, R, Tchoubrieva, E, Hannan, KM, Lin, JI, Sheppard, KE, Jastrzebski, K, Quinn, LM, Rothblum, LI, Pearson, RB, Hannan, RD & Sanij, E 2017, 'Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription' Gene, vol. 612, pp. 36-48. https://doi.org/10.1016/j.gene.2016.12.015

Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription. / Hung, Sandy; Lesmana, Analia; Peck, Abigail; Lee, Rachel; Tchoubrieva, Elly; Hannan, Katherine M; Lin, Jane I; Sheppard, Karen E; Jastrzebski, Katarzyna; Quinn, Leonie M; Rothblum, Lawrence I; Pearson, Richard B.; Hannan, Ross D.; Sanij, Elaine.

In: Gene, Vol. 612, 15.05.2017, p. 36-48.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Cell cycle and growth stimuli regulate different steps of RNA polymerase I transcription

AU - Hung, Sandy

AU - Lesmana, Analia

AU - Peck, Abigail

AU - Lee, Rachel

AU - Tchoubrieva, Elly

AU - Hannan, Katherine M

AU - Lin, Jane I

AU - Sheppard, Karen E

AU - Jastrzebski, Katarzyna

AU - Quinn, Leonie M

AU - Rothblum, Lawrence I

AU - Pearson, Richard B.

AU - Hannan, Ross D.

AU - Sanij, Elaine

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N2 - Transcription of the ribosomal RNA genes (rDNA) by RNA polymerase I (Pol I) is a major control step for ribosome synthesis and is tightly linked to cellular growth. However, the question of whether this process is modulated primarily at the level of transcription initiation or elongation is controversial. Studies in markedly different cell types have identified either initiation or elongation as the major control point. In this study, we have re-examined this question in NIH3T3 fibroblasts using a combination of metabolic labeling of the 47S rRNA, chromatin immunoprecipitation analysis of Pol I and overexpression of the transcription initiation factor Rrn3. Acute manipulation of growth factor levels altered rRNA synthesis rates over 8-fold without changing Pol I loading onto the rDNA. In fact, robust changes in Pol I loading were only observed under conditions where inhibition of rDNA transcription was associated with chronic serum starvation or cell cycle arrest. Overexpression of the transcription initiation factor Rrn3 increased loading of Pol I on the rDNA but failed to enhance rRNA synthesis in either serum starved, serum treated or G0/G1 arrested cells. Together these data suggest that transcription elongation is rate limiting for rRNA synthesis. We propose that transcription initiation is required for rDNA transcription in response to cell cycle cues, whereas elongation controls the dynamic range of rRNA synthesis output in response to acute growth factor modulation.

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