A genetic screen for terminator function in yeast identifies a role for a new functional domain in termination factor Nab3.

Nucleic Acids Research
Travis J LoyaDaniel Reines

Abstract

The yeast IMD2 gene encodes an enzyme involved in GTP synthesis. Its expression is controlled by guanine nucleotides through a set of alternate start sites and an intervening transcriptional terminator. In the off state, transcription results in a short non-coding RNA that starts upstream of the gene. Transcription terminates via the Nrd1-Nab3-Sen1 complex and is degraded by the nuclear exosome. Using a sensitive terminator read-through assay, we identified trans-acting Terminator Override (TOV) genes that operate this terminator. Four genes were identified: the RNA polymerase II phosphatase SSU72, the RNA polymerase II binding protein PCF11, the TRAMP subunit TRF4 and the hnRNP-like, NAB3. The TOV phenotype can be explained by the loss of function of these gene products as described in models in which termination and RNA degradation are coupled to the phosphorylation state of RNA polymerase II's repeat domain. The most interesting mutations were those found in NAB3, which led to the finding that the removal of merely three carboxy-terminal amino acids compromised Nab3's function. This region of previously unknown function is distant from the protein's well-known RNA binding and Nrd1 binding domains. Structural homology modelin...Continue Reading

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Citations

Jul 9, 2013·FEBS Letters·Ana M Rodríguez-TorresMaría A Freire-Picos
Feb 11, 2014·Molecular Cell·Mariona Nadal-RibellesFrancesc Posas
Jan 23, 2015·Prion·Thomas W O'RourkeDaniel Reines
May 10, 2015·Biochimie·Agnieszka TudekDomenico Libri
Oct 1, 2015·BioMed Research International·Xiao-Yang LiuYi-Peng Wang
Mar 10, 2015·Annual Review of Biochemistry·Karen M Arndt, Daniel Reines
Oct 23, 2012·Biochimica Et Biophysica Acta·Hannah E Mischo, Nick J Proudfoot
Mar 10, 2016·PloS One·Thomas W O'Rourke, Daniel Reines
Mar 19, 2020·PLoS Genetics·Jose F VictorinoAmber L Mosley
Mar 9, 2019·Wiley Interdisciplinary Reviews. RNA·Sarah A PeckAmber L Mosley

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Datasets Mentioned

BETA
YSC1021-669423

Methods Mentioned

BETA
PCR
FACS
fluorescence-activated cell sorting
flow cytometry
X-ray
fluorescence sorting
NMR

Software Mentioned

CLC Genomics Workbench
SCS2
SPRI works
FACSDiva
PHYRE
HHpred
BLAST
Illumina

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