Abstract
Internal initiation of translation, whereby ribosomes are directed to internal AUG codon independently of the 5' end of the mRNA, has been observed rarely in higher eucaryotes and has not been demonstrated in living yeast. We report here that starved yeast cells are capable of initiating translation of a dicistronic message internally. The studied element that functions as an internal ribosome entry site (IRES) is hardly functional or not functional at all in logarithmically growing cells. Moreover, during the logarithmic growth phase, this element seems to inhibit translation reinitiation when placed as an intercistronic spacer or to inhibit translation when placed in the 5'-untranslated region of a monocistronic message. Inhibition of translation is likely due to the putative strong secondary structure of the IRES that interferes with the cap-dependent scanning process. When cells exit the logarithmic growth phase, or when artificially starved for carbon source, translation of the IRES-containing messages is substantially induced. Our findings imply that the capacity to translate internally is a characteristic of starved rather than vegetatively growing yeast cells.
Highlights
Cells undergo terminal differentiation [10]
No internal ribosome entry site (IRES) has been shown to function in yeasts, despite the observations that the yeast cell-free system is capable of recognizing plant viral IRESes [14] as well as natural yeast leader sequences [15]
It is quite possible that the control of this dramatic strategic change in gene expression requires stationary phase (SP)-specific mechanisms for ensuring the synthesis of the small repertoire of proteins that are essential for life during starvation
Summary
Cells undergo terminal differentiation [10]. IRESes have been found to mediate the differential translation of Antenapedia and Ultrabithorax during Drosophila melanogaster development [6, 12]. During the logarithmic growth phase, the gal level in cells carrying the SIRES-containing construct was two orders of magnitude lower than that in cells carrying the control construct (log lanes, compare the right and left panels).
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