W. Zhou et al., Transcript leader regions of two Saccharomyces cerevisiae mRNAs contain internal ribosome entry sites that function in living cells, P NAS US, 98(4), 2001, pp. 1531-1536
Citations number
41
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
In higher eukaryotes, translation of some mRNAs occurs by internal initiati
on. It is not known, however, whether this mechanism is used to initiate th
e translation of any yeast mRNAs. In this report, we identify naturally occ
urring nucleotide sequences that function as internal ribosome entry sites
(IRESes) within the 5' leader sequences of Saccharomyces cerevisiae YAP1 an
d p150 mRNAs. When tested in the 5' untranslated regions of monocistronic r
eporter genes, both leader sequences enhanced translation efficiency in veg
etatively growing yeast cells. Moreover, when tested in the intercistronic
region of dicistronic mRNAs, both sequences were shown to contain IRESes th
at functioned in living cells. The activity of the p150 leader was much gre
ater than that of the YAP1 leader. The second cistron was not expressed in
control dicistronic constructs that lacked these sequences or contained the
5' leader sequence of the CLN3 mRNA in the intercistronic region. Further
analyses of the p150 IRES revealed that it contained several nonoverlapping
segments that were able independently to mediate internal initiation. Thes
e results suggested a modular composition for the p150 IRES that resembled
the composition of IRESes contained within some cellular mRNAs of higher eu
karyotes. Both YAP1 and p150 leaders contain several complementary sequence
matches to yeast 18S rRNA. The findings are discussed in terms of our unde
rstanding of internal initiation in higher eukaryotes.