PXD028496

PXD028496 is an original dataset announced via ProteomeXchange.

Title	Quantification of proline to serine mistranslation in a Saccharomyces cerevisiae strain expressing an inducible tRNA variant
Description	Altering the genetic code for applications in synthetic biology and genetic code expansion involves engineered tRNAs that incorporate amino acids that differ from what is defined by the “standard” genetic code. Since these engineered tRNA variants can be lethal due to proteotoxic stress, regulating their expression is necessary to achieve high levels of the resulting novel proteins. Mechanisms to positively regulate transcription with exogenous activator proteins like those often used to regulate RNA polymerase II (RNAP II) transcribed genes are not applicable to tRNAs as their expression by RNA polymerase III requires elements internal to the tRNA. Here, we show that tRNA expression is repressed by overlapping transcription from an adjacent RNAP II promoter. Regulating the expression of the RNAP II promoter allows inverse regulation of the tRNA. Placing either Gal4 or TetR-VP16 activated promoters downstream of a mistranslating tRNA serine variant that mis-incorporates serine at proline codons in Saccharomyces cerevisiae allows mistranslation at a level not otherwise possible because of the toxicity of the unregulated tRNA. Using mass spectrometry, we determine th frequency of mistranslation in both the induced and repressed conditions of the galactose inducible and tetracycline inducible systems.
HostingRepository	PRIDE
AnnounceDate	2021-11-03
AnnouncementXML	Submission_2021-11-03_03:40:41.109.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Matthew Berg
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	L-methionine sulfone; S-carboxamidomethyl-L-cysteine
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2021-09-16 00:00:53	ID requested
⏵ 1	2021-11-03 03:40:41	announced

Dataset with its publication pending

submitter keyword: Yeast, mistranslation, tRNA variants, inducible

Judit Villen
contact affiliation	Department of Genome Sciences, University of Washington, Seattle, USA
contact email	jvillen@uw.edu
lab head
Matthew Berg
contact affiliation	Department of Genome Sciences, University of Washington
contact email	mattberg@uw.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/11/PXD028496

PRIDE project URI

• PRIDE
  1. PXD028496
     1. Label: PRIDE project
     2. Name: Quantification of proline to serine mistranslation in a Saccharomyces cerevisiae strain expressing an inducible tRNA variant