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. 2005 Oct 21;310(5747):486-9.
doi: 10.1126/science.1115791. Epub 2005 Sep 1.

Movement of eukaryotic mRNAs between polysomes and cytoplasmic processing bodies

Muriel Brengues  1 , Daniela Teixeira, Roy Parker
Affiliations

Movement of eukaryotic mRNAs between polysomes and cytoplasmic processing bodies

VSports在线直播 - Muriel Brengues et al. Science. .

Abstract

Eukaryotic cells contain nontranslating messenger RNA concentrated in P-bodies, which are sites where the mRNA can be decapped and degraded VSports手机版. We present evidence that mRNA molecules within yeast P-bodies can also return to translation. First, inhibiting delivery of new mRNAs to P-bodies leads to their disassembly independent of mRNA decay. Second, P-bodies decline in a translation initiation-dependent manner during stress recovery. Third, reporter mRNAs concentrate in P-bodies when translation initiation is blocked and resume translation and exit P-bodies when translation is restored. Fourth, stationary phase yeast have large P-bodies containing mRNAs that reenter translation when growth resumes. The reciprocal movement of mRNAs between polysomes and P-bodies is likely to be important in the control of mRNA translation and degradation. Moreover, the presence of related proteins in P-bodies and maternal mRNA storage granules suggests this mechanism is widely adapted for mRNA storage. .

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Figures

Fig. 1
Fig. 1
mRNAs exit P-bodies in the absence of decapping. (A) Wild-type (WT) and dcp1Δ cells grown in yeast extract, peptone, and glucose (YPGlu) at 30°C (control) were treated with cycloheximide (CYH, 100 μg/ml) for 10 min. (B) Cells grown in YPGlu at 23°C were shifted to 37°C for 30 min followed by CYH treatment for 10 min.
Fig. 2
Fig. 2
Disassembly of P-bodies after translation restoration. WT cells expressing a GFP-tagged version of Dcp2p (B, G, and L), Dhh1p (C, H, and M), or co-expressing U1A-GFP with PGK1-U1A (D, I, and N) or MFA2P-U1A (E, J, and O), were grown in glucose containing medium at 30°C (Glu), shifted for 10 min without glucose (10 min −Glu), followed by readdition of glucose (5 min +Glu). Polysome profiles under each condition are shown (A, F, and K).
Fig. 3
Fig. 3
Translation initiation, but not decapping, is required for mRNAs to exit P-bodies. Cells were grown in glucose-containing medium at 23°C (Glu), shifted to 37°C for 10 min without glucose (10 min −Glu), followed by readdition of glucose (10 min +Glu). Polysome profiles under each condition are shown (a, f, and k). (A) WT, (B) dcp1-2, and (C) prt1-63 cells expressing a GFP-tagged version of Dcp2p (b, g, and l), Dhh1p (c, h, and m), or co-expressing the plasmids U1A-GFP and PGK1-U1A (d, i, and n), or U1A-GFP and MFA2P-U1A (e, j, and o) are shown.
Fig. 4
Fig. 4
Movement of mRNAs between a translating and a nontranslating pool. (A) WT cells co-expressing U1A-GFP with MFA2P-U1A or (B) expressing a plasmid containing MFA2P-U1A or MFA2pG under the control of a tetracycline promoter (Tet-Off MFA2P-U1A and Tet-Off MFA2pG) were grown at 30°C in synthetic complete (SC) medium plus glucose (Glu), shifted for 10 min to (A) SC with no glucose or (B) SC with no glucose containing doxycycline (10 min −Glu), followed by readdition of glucose (5 min +Glu). (A) Polysome profiles of the collected sucrose gradients are shown. Fraction 1 corresponds to the top of the gradient. (A and B) Northern blots for the indicated mRNA are shown.
Fig. 5
Fig. 5
mRNA present in P-bodies in stationary phase can return to translation. Cells expressing Tet-Off MFA2pG or co-expressing U1A-GFP with MFA2P-U1A were grown until stationary phase (SP). Then glucose, with or without doxycycline, was added to the culture (30 min +Glu). Polysome profiles and Northern blotting of cells expressing the plasmid Tet-Off MFA2pG are shown (top). Cells co-expressing U1A-GFP and MFA2P-U1A are visualized (bottom). In stationary phase, ~80% of the cells show large foci; after 30 min +Glu, ~20% of the cells have foci, and the remaining foci are considerably diminished.
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