RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output
How eukaryotic RNA polymerases switch from elongation to termination is unknown. Pol III subunits Rpc53 and Rpc37 (C53/37) form a heterodimer homologous to TFIIFβ/α. C53/37 promotes efficient termination and together with C11 also mediates pol III recycling in vitro. We previously developed Schizosa...
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Oxford University Press
2013
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3592421/ |
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pubmed-35924212013-03-08 RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output Rijal, Keshab Maraia, Richard J. Gene Regulation, Chromatin and Epigenetics How eukaryotic RNA polymerases switch from elongation to termination is unknown. Pol III subunits Rpc53 and Rpc37 (C53/37) form a heterodimer homologous to TFIIFβ/α. C53/37 promotes efficient termination and together with C11 also mediates pol III recycling in vitro. We previously developed Schizosaccharomyces pombe strains that report on two pol III termination activities: RNA oligo(U) 3′-end cleavage, and terminator readthrough. We randomly mutagenized C53 and C37 and isolated many C37 mutants with terminator readthrough but no comparable C53 mutants. The majority of C37 mutants have strong phenotypes with up to 40% readthrough and map to a C-terminal tract previously localized near Rpc2p in the pol III active center while a minority represent a distinct class with weaker phenotype, less readthrough and 3′-oligo(U) lengthening. Nascent pre-tRNAs released from a terminator by C37 mutants have shorter 3′-oligo(U) tracts than in cleavage-deficient C11 double mutants indicating RNA 3′-end cleavage during termination. We asked whether termination deficiency affects transcription output in the mutants in vivo both by monitoring intron-containing nascent transcript levels and 14C-uridine incorporation. Surprisingly, multiple termination mutants have no decrease in transcript output relative to controls. These data are discussed in context of current models of pol III transcription. Oxford University Press 2013-01 2012-10-23 /pmc/articles/PMC3592421/ /pubmed/23093604 http://dx.doi.org/10.1093/nar/gks985 Text en Published by Oxford University Press 2012. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Rijal, Keshab Maraia, Richard J. |
| spellingShingle |
Rijal, Keshab Maraia, Richard J. RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| author_facet |
Rijal, Keshab Maraia, Richard J. |
| author_sort |
Rijal, Keshab |
| title |
RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| title_short |
RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| title_full |
RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| title_fullStr |
RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| title_full_unstemmed |
RNA polymerase III mutants in TFIIFα-like C37 that cause terminator readthrough with no decrease in transcription output |
| title_sort |
rna polymerase iii mutants in tfiifα-like c37 that cause terminator readthrough with no decrease in transcription output |
| description |
How eukaryotic RNA polymerases switch from elongation to termination is unknown. Pol III subunits Rpc53 and Rpc37 (C53/37) form a heterodimer homologous to TFIIFβ/α. C53/37 promotes efficient termination and together with C11 also mediates pol III recycling in vitro. We previously developed Schizosaccharomyces pombe strains that report on two pol III termination activities: RNA oligo(U) 3′-end cleavage, and terminator readthrough. We randomly mutagenized C53 and C37 and isolated many C37 mutants with terminator readthrough but no comparable C53 mutants. The majority of C37 mutants have strong phenotypes with up to 40% readthrough and map to a C-terminal tract previously localized near Rpc2p in the pol III active center while a minority represent a distinct class with weaker phenotype, less readthrough and 3′-oligo(U) lengthening. Nascent pre-tRNAs released from a terminator by C37 mutants have shorter 3′-oligo(U) tracts than in cleavage-deficient C11 double mutants indicating RNA 3′-end cleavage during termination. We asked whether termination deficiency affects transcription output in the mutants in vivo both by monitoring intron-containing nascent transcript levels and 14C-uridine incorporation. Surprisingly, multiple termination mutants have no decrease in transcript output relative to controls. These data are discussed in context of current models of pol III transcription. |
| publisher |
Oxford University Press |
| publishDate |
2013 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3592421/ |
| _version_ |
1611960833363935232 |