Monitoring Monomer-Specific Acyl-tRNA Levels in Cells with PARTI
Introduction
Study Focus: The study presents a new assay named PARTI (3-Prime Adenosine-Retaining Aminoacyl-tRNA Isolation) to monitor acylation states of tRNAs in cells.
Relevance: Understanding the cellular incorporation of non-amino acid monomers into proteins.
Authors and Institutions
Authors: Meghan A. Pressimone, Carly K. Schissel, Isabella H. Goss, Cameron V. Swenson, Alanna Schepartz
Institutions:
University of California, Berkeley (Departments of Molecular and Cellular Biology, and Chemistry)
Institute for Quantitative Biosciences (QB3)
Chan Zuckerberg Biohub
ARC Institute
Methodology
Assay Description:
PARTI relies on high-resolution mass spectrometry to identify acyl-adenosine species released by RNase A cleavage of isolated cellular tRNA.
Provides a direct report on the acylation state of a user-chosen tRNA.
Key Observations and Applications
Applications:
Investigating selectivity of translation with 2-hydroxy acid enantiomers.
Examining activity of PylRS variants for benzyl derivatives of malonic acid.
Addressing the inability of N-Me amino acids to function as ribosome substrates.
Findings: Direct evidence for cellular production of 2,3-diacylated tRNA.
Significance
Benefits: The simplicity of the PARTI workflow aids in studying and improving non-amino acid monomer incorporation into proteins.
Competing Interest Statement
Disclosure: Authors declared no competing interests.