Active Center Control of Termination by RNA Polymerase III and tRNA Gene Transcription Levels <i>In Vivo</i>

The ability of RNA polymerase (RNAP) III to efficiently recycle from termination to reinitiation is critical for abundant tRNA production during cellular proliferation, development and cancer. Yet understanding of the unique termination mechanisms used by RNAP III is incomplete, as is its link to high transcription output. We used two tRNA-mediated suppression systems to screen for Rpc1 mutants with gain- and loss- of termination phenotypes in S. pombe. 122 point mutation mutants were mapped to a recently solved 3.9 Å structure of yeast RNAP III elongation complex (EC); they cluster in the active center bridge helix and trigger loop, as well as the pore and funnel, the latter of which indicate involvement of the RNA cleavage domain of the C11 subunit in termination. Purified RNAP III from a readthrough (RT) mutant exhibits increased elongation rate. The data strongly support a kinetic coupling model in which elongation rate is inversely related to termination efficiency. The mutants exhibit good correlations of terminator RT in vitro and in vivo, and surprisingly, amounts of transcription in vivo. Because assessing in vivo transcription can be confounded by various parameters, we used a tRNA reporter with a processing defect and a strong terminator. By ruling out differences in RNA decay rates, the data indicate that mutants with the RT phenotype synthesize more RNA than wild type cells, and than can be accounted for by their increased elongation rate. Finally, increased activity by the mutants appears unrelated to the RNAP III repressor, Maf1. The results show that the mobile elements of the RNAP III active center, including C11, are key determinants of termination, and that some of the mutations activate RNAP III for overall transcription. Similar mutations in spontaneous cancer suggest this as an unforeseen mechanism of RNAP III activation in disease.

Tags
Data and Resources
To access the resources you must log in

This item has no data

Identity

Description: The Identity category includes attributes that support the identification of the resource.

Field Value
PID https://www.doi.org/10.1371/journal.pgen.1006253
URL https://figshare.com/articles/Active_Center_Control_of_Termination_by_RNA_Polymerase_III_and_tRNA_Gene_Transcription_Levels_i_In_Vivo_i_/3578802
URL http://dx.doi.org/10.1371/journal.pgen.1006253
Access Modality

Description: The Access Modality category includes attributes that report the modality of exploitation of the resource.

Field Value
Access Right Open Access
Attribution

Description: Authorships and contributors

Field Value
Author Rijal, Keshab
Author J. Maraia, Richard
Publishing

Description: Attributes about the publishing venue (e.g. journal) and deposit location (e.g. repository)

Field Value
Collected From figshare
Hosted By figshare
Publication Date 2016-01-01
Publisher Figshare
Additional Info
Field Value
Language UNKNOWN
Resource Type Dataset
keyword 3.9 Å structure
system:type dataset
Management Info
Field Value
Source https://science-innovation-policy.openaire.eu/search/dataset?datasetId=r37980778c78::74f1c44807e3208c781a4825008aa34c
Author jsonws_user
Last Updated 12 January 2021, 15:25 (CET)
Created 12 January 2021, 15:25 (CET)