Analysis of Human Endogenous mRNA Deadenylation Complexes by High-Resolution Gel Electrophoresis

Chung Te Chang*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

In eukaryotic cells, poly(A) tails stabilize mRNA molecules and play a pivotal role in enhancing translational efficiency. Consequently, the enzymatic shortening of these poly(A) tails by deadenylase enzymes has a critical role in post-transcriptional gene regulation. However, deadenylases are usually large, multisubunit, and multifunctional complexes, which complicates their biochemical analysis. This chapter presents a methodology for isolating human deadenylation complexes from endogenous sources and conducting an in vitro deadenylation assay to examine their enzymatic activity. The reactions involving fluorescently labeled synthetic polyadenylated RNAs are subsequently analyzed using high-resolution denaturing polyacrylamide gel electrophoresis.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages47-54
Number of pages8
DOIs
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
Volume2723
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CCR4-NOT
  • Deadenylase
  • Poly(A) tail
  • Post-transcriptional regulation

Fingerprint

Dive into the research topics of 'Analysis of Human Endogenous mRNA Deadenylation Complexes by High-Resolution Gel Electrophoresis'. Together they form a unique fingerprint.

Cite this