Interaction of eukaryotic initiation factor eIF-4B with a picornavirus internal translation initiation site.
We studied the interaction of cellular proteins with the internal ribosome entry site (IRES) of foot-and-mouth disease virus by UV cross-linking and observed specific binding of a 80-kDa protein contained in cytosolic HeLa cell extract and in rabbit reticulocyte lysate. Binding of the protein was dependent on the presence of ATP. Immunoprecipitation with eIF-4B antiserum revealed that the protein is identical to the initiation factor eIF-4B. Deletions in the 3' part, but not in the 5' part, of the IRES interfered with UV cross-linking, indicating that the binding site of eIF-4B is located close to the end of the element. Attempts to separate ribosome-associated from non-ribosome-associated protein fractions of cytosolic cell extracts led to the loss of cross-linking activity. This finding suggests that additional protein factors contribute to this interaction of eIF-4B with the IRES of foot-and-mouth disease virus.
Meyer
K
K
Biochemisches Institut am Klinikum, Justus-Liebig-Universität Giessen, Germany.
Petersen
A
A
Niepmann
M
M
Beck
E
E
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D005537:Aphthovirus / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003432:Cross-Linking Reagents; D039642:Eukaryotic Initiation Factors; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010448:Peptide Initiation Factors / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D012270:Ribosomes / Q000378:metabolism; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D039642:Eukaryotic Initiation Factors; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D012367:RNA, Viral; C031799:eIF-4B; D000255:Adenosine Triphosphate