Publications
2018
Gross L, Schaeffer L, Alghoul F, Hayek H, Allmang C, Eriani G, Martin F
Tracking the m7G-cap during translation initiation by crosslinking methods Journal Article
In: Methods, vol. 137, pp. 3-10, 2018, ISBN: 29307728.
Abstract | Links | BibTeX | Tags: Cap-dependent translation Chemical crosslinking Histone H4 mRNA Ribosome UV crosslinking eIF4E, ERIANI, Unité ARN
@article{,
title = {Tracking the m^{7}G-cap during translation initiation by crosslinking methods},
author = {L Gross and L Schaeffer and F Alghoul and H Hayek and C Allmang and G Eriani and F Martin},
url = {https://www.ncbi.nlm.nih.gov/pubmed/29307728?dopt=Abstract},
doi = {10.1016/j.ymeth.2017.12.019},
isbn = {29307728},
year = {2018},
date = {2018-01-01},
journal = {Methods},
volume = {137},
pages = {3-10},
abstract = {In eukaryotes, cap-dependent translation initiation is a sophisticated process that requires numerous trans-acting factors, the eukaryotic Initiation Factors (eIFs). Their main function is to assist the ribosome for accurate AUG start codon recognition. The whole process requires a 5'-3' scanning step and is therefore highly dynamic. Therefore translation requires a complex interplay between eIFs through assembly/release cycles. Here, we describe an original approach to assess the dynamic features of translation initiation. The principle is to use the m7Gcap located at the 5' extremity of mRNAs as a tracker to monitor RNA and protein components that are in its vicinity. Cap-binding molecules are trapped by chemical and UV crosslinking. The combination of cap crosslinking methods in cell-free translation systems with the use of specific translation inhibitors for different steps such as edeine, GMP-PNP or cycloheximide allowed assessing the cap fate during eukaryotic translation. Here, we followed the position of the cap in the histone H4 mRNA and the cap binding proteins during H4 mRNA translation.},
keywords = {Cap-dependent translation Chemical crosslinking Histone H4 mRNA Ribosome UV crosslinking eIF4E, ERIANI, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
In eukaryotes, cap-dependent translation initiation is a sophisticated process that requires numerous trans-acting factors, the eukaryotic Initiation Factors (eIFs). Their main function is to assist the ribosome for accurate AUG start codon recognition. The whole process requires a 5'-3' scanning step and is therefore highly dynamic. Therefore translation requires a complex interplay between eIFs through assembly/release cycles. Here, we describe an original approach to assess the dynamic features of translation initiation. The principle is to use the m7Gcap located at the 5' extremity of mRNAs as a tracker to monitor RNA and protein components that are in its vicinity. Cap-binding molecules are trapped by chemical and UV crosslinking. The combination of cap crosslinking methods in cell-free translation systems with the use of specific translation inhibitors for different steps such as edeine, GMP-PNP or cycloheximide allowed assessing the cap fate during eukaryotic translation. Here, we followed the position of the cap in the histone H4 mRNA and the cap binding proteins during H4 mRNA translation.