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tRNA Activation

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Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...

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Updated: May 31, 2026

Transcription Start Site Mapping Using Super-low Input Carrier-CAGE

Transcription Start Site Mapping Using Super-low Input Carrier-CAGE

Published on: June 26, 2019

Structural insights to how mammalian capping enzyme reads the CTD code.

Agnidipta Ghosh¹, Stewart Shuman, Christopher D Lima

¹Structural Biology Program, Sloan-Kettering Institute, New York, NY 10065, USA.

|June 21, 2011

Summary

The crystal structure reveals how mammalian capping enzymes bind to phosphorylated RNA polymerase II CTD, crucial for mRNA capping. This interaction differs from yeast enzymes, showing evolved strategies for reading the CTD code.

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An Integrated Approach for Microprotein Identification and Sequence Analysis

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Published on: July 12, 2022

Area of Science:

Molecular Biology
Biochemistry
Structural Biology

Background:

Efficient 5' mRNA cap formation requires interaction between phosphorylated RNA polymerase II CTD and capping enzymes.
The 5' mRNA cap is the initial modification of nascent mRNA.

Purpose of the Study:

To determine the crystal structure of the RNA guanylyltransferase component of mammalian capping enzyme (Mce) bound to a CTD phosphopeptide.
To understand the molecular basis of CTD recognition by Mce and its functional implications.

Main Methods:

X-ray crystallography to obtain the Mce-CTD phosphopeptide complex structure.
Structure-guided mutational analysis to probe the Mce-CTD interface.
In vivo functional assays to assess Mce activity.

Main Results:

The CTD adopts an extended β-like conformation, docking Tyr1 and Ser5-PO(4) onto the Mce nucleotidyltransferase domain.
The Mce-CTD interface was identified as a key determinant of CTD binding, guanylyltransferase activity stimulation, and Mce function in vivo.
The CTD binding site on mammalian capping enzyme is distinct from that of yeast capping enzyme.

Conclusions:

Mammalian capping enzymes utilize a distinct structural strategy to bind the phosphorylated CTD compared to yeast enzymes.
Understanding these differences provides insights into the evolution of mRNA capping machinery.
The Mce-CTD interaction is a tunable regulatory point for mRNA capping.