Effects of poly(A)-binding protein on the interactions of translation initiation factor eIF4F and eIF4F.4B with internal ribosome entry site (IRES) of tobacco etch virus RNA.
In wheat germ, the interaction between poly(A)-binding protein and eukaryotic initiation factor eIF 4G increases the affinity of eIF4E for the cap by 20-40-fold. Recent findings that wheat germ eIF4G is required for interaction with the IRES, pseudoknot 1 (PK1), of tobacco etch virus to promote cap-independent translation led us to investigate the effects of PABP on the interaction of eIF4F with PK1. The fluorescence anisotropy data showed addition of PABP to eIF4F increased the binding affinity approximately 2.0-fold for PK1 RNA as compared with eIF4F alone. Addition of both PABP and eIF4B to eIF4F enhance binding affinity to PK1 about 4-fold, showing an additive effect rather than the large increase in affinity shown for cap binding. The van't Hoff analyses showed that PK1 RNA binding to eIF4F, eIF4F.PABP, eIF4F.4B and eIF4F.4B.PABP is enthalpy-driven and entropy-favorable. PABP and eIF4B decreased the entropic contribution 65% for binding of PK1 RNA to eIF4F. The lowering of entropy for the formation of eIF4F.4B.PABP-PK1 complex suggested reduced hydrophobic interactions for complex formation. Overall, these results demonstrate the first direct effect of PABP on the interaction of eIF4F and eIF4F.4B with PK1 RNA.
Khan
Mateen A
MA
Department of Chemistry, Hunter College and the Graduate Center of the City University of New York, New York, NY 10065, USA.
Yumak
Hasan
H
Gallie
Daniel R
DR
Goss
Dixie J
DJ
D001665:Binding Sites / Q000187:drug effects; D039562:Eukaryotic Initiation Factor-4F / Q000378:metabolism*; D039642:Eukaryotic Initiation Factors / Q000378:metabolism*; D005454:Fluorescence Polarization; D057927:Hydrophobic and Hydrophilic Interactions; D010849:Picornaviridae / Q000235:genetics*; D039102:Poly(A)-Binding Proteins / Q000494:pharmacology / Q000502:physiology*; D011485:Protein Binding / Q000187:drug effects; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D013696:Temperature; D014026:Nicotiana / Q000821:virology; D014908:Triticum / Q000378:metabolism
D039562:Eukaryotic Initiation Factor-4F; D039642:Eukaryotic Initiation Factors; D039102:Poly(A)-Binding Proteins; D012367:RNA, Viral; C031799:eIF-4B