Adapted formaldehyde gradient cross-linking protocol implicates human eIF3d and eIF3c, k and I subunits in the 43S and 48S pre-initiation complex assembly, respectively

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F20%3A00524740" target="_blank" >RIV/61388971:_____/20:00524740 - isvavai.cz</a>

Result on the web

<a href="https://academic.oup.com/nar/article/48/4/1969/5682903" target="_blank" >https://academic.oup.com/nar/article/48/4/1969/5682903</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1093/nar/gkz1185" target="_blank" >10.1093/nar/gkz1185</a>

Result language

angličtina

Original language name

Adapted formaldehyde gradient cross-linking protocol implicates human eIF3d and eIF3c, k and I subunits in the 43S and 48S pre-initiation complex assembly, respectively

Original language description

One of the key roles of the 12-subunit eukaryotic translation initiation factor 3 (eIF3) is to promote the formation of the 43S and 48S pre-initiation complexes (PICs). However, particular contributions of its individual subunits to these two critical initiation reactions remained obscure. Here, we adapted formaldehyde gradient cross-linking protocol to translation studies and investigated the efficiency of the 43S and 48S PIC assembly in knockdowns of individual subunits of human eIF3 known to produce various partial subcomplexes. We revealed that eIF3d constitutes an important intermolecular bridge between eIF3 and the 40S subunit as its elimination from the eIF3 holocomplex severely compromised the 43S PIC assembly. Similarly, subunits elF3a, c and e were found to represent an important binding force driving eIF3 binding to the 40S subunit. In addition, we demonstrated that eIF3c, and eIF3k and I subunits alter the efficiency of mRNA recruitment to 43S PICs in an opposite manner. Whereas the eIF3c knockdown reduces it, downregulation of eIF3k or eIF3I increases mRNA recruitment, suggesting that the latter subunits possess a regulatory potential. Altogether this study provides new insights into the role of human eIF3 in the initial assembly steps of the translational machinery.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10606 - Microbiology

Project

<a href="/en/project/GA17-06238S" target="_blank" >GA17-06238S: In-depth analysis of the function and regulatory potential of individual subunits of human translation initiation factor 3 and their subcomplexes.</a><br>

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2020

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Nucleic Acids Research

ISSN

0305-1048

e-ISSN

—

Volume of the periodical

48

Issue of the periodical within the volume

4

Country of publishing house

GB - UNITED KINGDOM

Number of pages

16

Pages from-to

1969-1984

UT code for WoS article

000525957000030

EID of the result in the Scopus database

2-s2.0-85081103124