TY - JOUR
T1 - Dynamic recognition of the mRNA cap by Saccharomyces cerevisiae eIF4E
AU - O'Leary, Seán E.
AU - Petrov, Alexey
AU - Chen, Jin
AU - Puglisi, Joseph D.
N1 - Funding Information:
This work was supported by NIH grants GM073999 and GM099687 (to J.D.P.).
PY - 2013/12/3
Y1 - 2013/12/3
N2 - Recognition of the mRNA 5′ m7G(5′)ppp(5′)N cap is key to translation initiation for most eukaryotic mRNAs. The cap is bound by the eIF4F complex, consisting of a cap-binding protein (eIF4E), a "scaffold" protein (eIF4G), and an RNA helicase (eIF4A). As a central early step in initiation, regulation of eIF4F is crucial for cellular viability. Although the structure and function of eIF4E have been defined, a dynamic mechanistic picture of its activity at the molecular level in the eIF4F×mRNA complex is still unavailable. Here, using single-molecule fluorescence, we measured the effects of Saccharomyces cerevisiae eIF4F factors, mRNA secondary structure, and the poly(A)-binding protein Pab1p on eIF4E-mRNA binding dynamics. Our data provide an integrated picture of how eIF4G and mRNA structure modulate eIF4E-mRNA interaction, and uncover an eIF4G- and poly(A)-independent activity of poly(A)-binding protein that prolongs the eIF4E×mRNA complex lifetime.
AB - Recognition of the mRNA 5′ m7G(5′)ppp(5′)N cap is key to translation initiation for most eukaryotic mRNAs. The cap is bound by the eIF4F complex, consisting of a cap-binding protein (eIF4E), a "scaffold" protein (eIF4G), and an RNA helicase (eIF4A). As a central early step in initiation, regulation of eIF4F is crucial for cellular viability. Although the structure and function of eIF4E have been defined, a dynamic mechanistic picture of its activity at the molecular level in the eIF4F×mRNA complex is still unavailable. Here, using single-molecule fluorescence, we measured the effects of Saccharomyces cerevisiae eIF4F factors, mRNA secondary structure, and the poly(A)-binding protein Pab1p on eIF4E-mRNA binding dynamics. Our data provide an integrated picture of how eIF4G and mRNA structure modulate eIF4E-mRNA interaction, and uncover an eIF4G- and poly(A)-independent activity of poly(A)-binding protein that prolongs the eIF4E×mRNA complex lifetime.
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U2 - 10.1016/j.str.2013.09.016
DO - 10.1016/j.str.2013.09.016
M3 - Article
C2 - 24183571
AN - SCOPUS:84889609770
SN - 0969-2126
VL - 21
SP - 2197
EP - 2207
JO - Structure
JF - Structure
IS - 12
ER -