An absolute requirement for the 5' cap structure for mRNA translation in sea urchin eggs.
Translation of a variety of RNAs was studied in a cell-free translation system derived from sea urchin eggs. While RNAs such as globin or tobacco mosaic virus are efficiently translated, viral RNAs which do not contain the 5' cap structure, such as cow pea mosaic virus (CPMV) and poliovirus, are not translated. Mixing experiments with reticulocyte lysates indicated that the lack of translation of uncapped viral RNAs is not due to the presence of a potent inhibitor or the absence of an activating agent. RNA competition experiments between capped and uncapped RNAs indicated that uncapped RNAs do not interact with the sea urchin egg initiation machinery. Proteolytic removal of the 5' viral protein did not allow the translation of CPMV RNA. However, chemical decapping of vesicular stomatitis virus mRNA completely inhibited the translation of this mRNA in the sea urchin cell-free system. We conclude that the sea urchin egg lacks the initiation pathway used to initiate uncapped mRNAs in mammalian cells and thus has an absolute requirement for the 5' cap structure for initiation. In addition we discuss the implications of these findings for the control of protein synthesis after fertilization of the sea urchin egg.
D000818:Animals; D002474:Cell-Free System; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D066298:In Vitro Techniques; D010063:Ovum / Q000378:metabolism*; D014176:Protein Biosynthesis; D012315:RNA Caps / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D012156:Reticulocytes / Q000378:metabolism; D012617:Sea Urchins
D012315:RNA Caps; D012333:RNA, Messenger; D012367:RNA, Viral