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

Kód výsledku v 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>

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/GA17-06238S" target="_blank" >GA17-06238S: Detailní analýza funkcí a regulačního potenciálu jednotlivých podjednotek lidského translačního iniciačního faktoru 3 a jejich dílčích pod-komplexů.</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2020

Kód důvěrnosti údajů

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

Název periodika

Nucleic Acids Research

ISSN

0305-1048

e-ISSN

—

Svazek periodika

48

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

16

Strana od-do

1969-1984

Kód UT WoS článku

000525957000030

EID výsledku v databázi Scopus

2-s2.0-85081103124