The decapping enzyme Dcp1 participates in translation termination through its interaction with the release factor eRF3 in budding yeast
Section snippets
Materials and methods
Yeast strains. All yeast strains used in this study are listed in Table 1. The yeast cells were grown in standard culture media and transformed with DNA by the lithium acetate method. Disruption of yeast genes was performed using PCR-based methods [9], and the disruption was confirmed by the phenotypic analysis and/or polymerase chain reactions (PCRs) with primers specific for the genes. Epitope tagging of yeast eRF3 (SUP35) was performed by the one-step method described by [10]. The
The decapping enzyme Dcp1p interacts with the eukaryotic release factor eRF3p/Sup35p
To elucidate the relationship between the decapping reaction and the translation termination reaction, we examined whether the decapping enzyme Dcp1p interacts with the release factor eRF3p in yeast. For the analysis, we introduced the plasmid expressing FLAG-tagged Dcp1p into wild-type cells whose chromosomal copy of SUP35 (encoding yeast eRF3) was tagged with Myc at its C terminus. As shown in Fig. 1, immunoprecipitation assay revealed that FLAG-Dcp1p interacts with eRF3p-Myc (the lower
Discussion
In S. cerevisiae, overexpression of both eRF1p and eRF3p enhances the translation termination at all three stop codons [15], indicating that the eRF1p–eRF3p complex acts as a key player at the ribosomal A site in the termination reaction. We previously reported that the GTP-bound form of eRF3p is required for the interaction with eRF1p to stimulate the termination reaction [3]. As shown in Fig. 5, eRF3 consists of an EF1α-like (GTP-binding) C-domain and a unique short N-domain [16]. The
Acknowledgments
This work was supported in part by research grants from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of the Japanese Government, Japan Society for the Promotion of Science (JSPS), the Mitsubishi Foundation, and the Uehara Memorial Foundation.
References (23)
- et al.
Translation termination factor eRF3 mediates mRNA decay through the regulation of deadenylation
J. Biol. Chem.
(2003) - et al.
The GTP-binding release factor eRF3 as a key mediator coupling translation termination to mRNA decay
J. Biol. Chem.
(2004) - et al.
A novel role of the mammalian GSPT/eRF3 associating with poly(A)-binding protein in Cap/Poly(A)-dependent translation
J. Biol. Chem.
(2002) - et al.
The eukaryotic polypeptide chain releasing factor (eRF3/GSPT) carrying the translation termination signal to the 3′-Poly(A) tail of mRNA. Direct association of erf3/GSPT with polyadenylate-binding protein
J. Biol. Chem.
(1999) - et al.
Differential effects of translational inhibition in cis and in trans on the decay of the unstable yeast MFA2 mRNA
J. Biol. Chem.
(1994) - et al.
General translational repression by activators of mRNA decapping
Cell
(2005) - et al.
Elevated recombination rates in transcriptionally active DNA
Cell
(1989) - et al.
The enzymes and control of eukaryotic mRNA turnover
Nat. Struct. Mol. Biol.
(2004) - et al.
Eukaryotic mRNA decapping
Annu. Rev. Biochem.
(2004) - et al.
Decapping reaction of mRNA requires Dcp1 in fission yeast: its characterization in different species from yeast to human
J. Biochem. (Tokyo)
(2004)
Identification of a human decapping complex associated with hUpf proteins in nonsense-mediated decay
Mol. Cell Biol.
Cited by (4)
The fate of the messenger is pre-determined: A new model for regulation of gene expression
2013, Biochimica et Biophysica Acta - Gene Regulatory MechanismsCitation Excerpt :eRF3 has been shown previously to mediate poly(A) shortening and mRNA decay in a manner coupled to translation termination [47]. Furthermore, yeast eRF3 directly interacts with the decapping protein Dcp1p [48], thus linking translation termination to decapping. Significantly, eRF3 and human PAN2/3 deadenylase compete for their binding to the poly(A) binding protein, PABPC1.
Mutants of the pafl complex alter phenotypic expression of the yeast prion [PSI+]
2009, Molecular Biology of the Cell
- 1
These authors contributed equally to this work.
- 2
Present address: Laboratory of Chromosome Dynamics, Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Tokyo 113-0032, Japan.
- 3
Present address: Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Nagoya 467-8603, Japan.