The immediate upstream region of the 5'-UTR from the AUG start codon has a pronounced effect on the translational efficiency in Arabidopsis thaliana.

The nucleotide sequence around the translational initiation site is an important cis-acting element for post-transcriptional regulation. However, it has not been fully understood how the sequence context at the 5'-untranslated region (5'-UTR) affects the translational efficiency of individual mRNAs. In this study, we provide evidence that the 5'-UTRs of Arabidopsis genes showing a great difference in the nucleotide sequence vary greatly in translational efficiency with more than a 200-fold difference. Of the four types of nucleotides, the A residue was the most favourable nucleotide from positions -1 to -21 of the 5'-UTRs in Arabidopsis genes. In particular, the A residue in the 5'-UTR from positions -1 to -5 was required for a high-level translational efficiency. In contrast, the T residue in the 5'-UTR from positions -1 to -5 was the least favourable nucleotide in translational efficiency. Furthermore, the effect of the sequence context in the -1 to -21 region of the 5'-UTR was conserved in different plant species. Based on these observations, we propose that the sequence context immediately upstream of the AUG initiation codon plays a crucial role in determining the translational efficiency of plant genes.

Expression of wee1 and its related cell cycle components in mouse early stage follicles.

Wee1 is a kinase regulator of the M-phase promoting factor (a complex of cdc2 and cyclin B1). The present study was performed to determine the role(s) of wee1 in the early stages of mouse ovarian follicles. Expression of wee1 and the correlated cell cycle components, namely cdc2, cyclin B1, and cdc25C, was evaluated by immunohistochemistry. In addition, expression of Tyr15-phosphorylated cdc2 (cdc2-p) was also examined to determine whether wee1 kinase phosphorylates cdc2. Each component except cdc25C was found in the oocyte cytoplasm at all follicular stages, while cdc25C was not detected in primordial follicles. It was found primarily in ovarian interstitial cells and to a small extent in granulosa cells of the developing secondary follicles. To further confirm the expression of cell cycle components in the primordial follicular oocytes, day 1 ovaries were enzymatically and mechanically dissociated, then oocytes were isolated from somatic cells including pre-granulosa cells, and we confirmed that cdc2-p was expressed in oocytes of primordial follicles. The results of the present study led to the conclusion that wee1, without the counteracting cdc25C, would cause meiotic arrest of oocytes by inhibitory phosphorylation of cdc2. Expression of all these proteins in the granulosa cells of growing follicles may regulate granulosa cell mitosis concurrently with the growth of oocytes and follicles.

Oocyte-selective expression of MT transposon-like element, clone MTi7 and its role in oocyte maturation and embryo development.

Previously, we found MT transposon-like element, clone MTi7 (MTi7) is highly expressed in the mouse ovary. Here, we show that the MTi7 is expressed in the oocyte from the primordial to the preovulatory follicles. For RNA interference (RNAi), double stranded RNAs (dsRNAs) were prepared for MTi7 and c-mos, a control gene with known functions. Each dsRNA was microinjected into germinal vesicle (GV) stage oocytes or zygotes with pronuclei (PN), after which developmental changes, mRNA expression, and nuclear and microtubular organization were analyzed. We found a 43.4-53% GV arrest in the microinjected oocytes with a concomitant decrease in targeted mRNA expression. In MTi7 dsRNA-injected early and late PN zygotes, a 92.9% 1-cell arrest and 76.9% 2-cell arrest were observed, respectively. This is the first report of an oocyte-selective expression of MTi7 mRNA, and our results strongly suggest that MTi7 involved in the nuclear membrane breakdown during oocyte maturation and embryo development.