PKA-dependent Binding of mRNA to the Mitochondrial AKAP121 Protein

Ginsberg, Michael D.; Feliciello, Antonio; Jones, Jeffrey K.; Avvedimento, Enrico V.; Gottesman, Max E.

doi:10.1016/s0022-2836(03)00173-6

Cited by 94 publications

(93 citation statements)

References 41 publications

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“…Approximately half of the mRNAs encoding mitochondrial proteins are found in the vicinity of yeast mitochondria, suggesting a link between mRNA localization and protein translocation (6). A few studies directly show that mitochondrial targeting of mRNA can improve protein import and rescue respiratory defects (27)(28)(29). Here we show that differences in VDAC3 mRNA targeting impact mitochondria morphology and size but do not cause differences in VDAC3 protein import efficiency.…”

Section: Discussionmentioning

confidence: 52%

Differential targeting of VDAC3 mRNA isoforms influences mitochondria morphology

Michaud

Ubrig

Filleur

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Intracellular targeting of mRNAs has recently emerged as a prevalent mechanism to control protein localization. For mitochondria, a cotranslational model of protein import is now proposed in parallel to the conventional posttranslational model, and mitochondrial targeting of mRNAs has been demonstrated in various organisms. Voltage-dependent anion channels (VDACs) are the most abundant proteins in the outer mitochondrial membrane and the major transport pathway for numerous metabolites. Four nucleus-encoded VDACs have been identified in Arabidopsis thaliana. Alternative cleavage and polyadenylation generate two VDAC3 mRNA isoforms differing by their 3′ UTR. By using quantitative RT-PCR and in vivo mRNA visualization approaches, the two mRNA variants were shown differentially associated with mitochondria. The longest mRNA presents a 3′ extension named alternative UTR (aUTR) that is necessary and sufficient to target VDAC3 mRNA to the mitochondrial surface. Moreover, aUTR is sufficient for the mitochondrial targeting of a reporter transcript, and can be used as a tool to target an unrelated mRNA to the mitochondrial surface. Finally, VDAC3-aUTR mRNA variant impacts mitochondria morphology and size, demonstrating the role of mRNA targeting in mitochondria biogenesis.mRNA localization | mRNA sorting | green RNA | porin | plant E ukaryotic cells are characterized by compartmentalization of functions in complex organelles, and cell activity depends on synthesis and precise localization of proteins. The mechanisms of protein sorting are the subject of intense research, and models are often reevaluated (1). The earliest models for protein localization were based on the presence of targeting signals in the polypeptide sequences. However, the targeting of mRNAs to various subcellular regions has recently emerged as an essential regulatory step for protein localization (2).Mitochondria contain hundreds of proteins, but only a few are encoded by the mitochondrial genome. The other proteins are nucleus-encoded and imported into the organelle. Protein import into mitochondria is controlled by peptide sequences that are recognized by mitochondrial receptors (3, 4). In parallel with this posttranslational model, a cotranslational model is now emerging (2, 5). A microarray analysis in Saccharomyces cerevisiae has shown that approximately half of the mRNAs encoding mitochondrial proteins are found in the vicinity of mitochondria (6). Mitochondrial targeting of mRNA was also demonstrated in mammals and plants (7,8).Mechanisms of mitochondrial targeting of mRNAs were essentially studied in yeast. In this species, three factors have been clearly identified to influence mRNA targeting to the mitochondrial surface. (i) Targeting of many but not all mRNAs appears dependent on translation (9, 10). (ii) The RNA-binding protein Puf3p is essential for mitochondrial targeting of a subset of mRNAs (9). (iii) The translocase of the outer mitochondrial membrane (TOM complex) is also involved in mRNA binding (10-12). In addition, some mRNAs are...

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Section: Discussionmentioning

confidence: 52%

Differential targeting of VDAC3 mRNA isoforms influences mitochondria morphology

Michaud

Ubrig

Filleur

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Immunoreactive bands were detected with the ECL kit (Amersham). Discrimination among the distinct types of extracts was ensured by running parallel western blots with the endogenous protein tubulin (localized to the PM), Mn-SOD (localized to the MM), VDAC and AKAP121 (localized to the OMM) (Cardone et al, 2004;Ginsberg et al, 2003;Liu et al, 2012), and COX-IV (localized to the IMM) (Mattei et al, 2011).…”

Section: Extraction Of Brain Mitochondria By the Percoll Gradient Methodsmentioning

confidence: 99%

NCX3 regulates mitochondrial calcium handling through AKAP121-anchored signaling complex and prevents hypoxia-induced cell death

Scorziello¹,

Savoia²,

Sisalli³

et al. 2013

Journal of Cell Science

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SummaryThe mitochondrial influx and efflux of Ca 2+ play a relevant role in cytosolic and mitochondrial Ca 2+ homeostasis, and contribute to the regulation of mitochondrial functions in neurons. The mitochondrial Na + /Ca 2+ exchanger, which was first postulated in 1974, has been primarily investigated only from a functional point of view, and its identity and localization in the mitochondria have been a matter of debate over the past three decades. Recently, a Li + -dependent Na + /Ca 2+ exchanger extruding Ca 2+ from the matrix has been found in the inner mitochondrial membrane of neuronal cells. However, evidence has been provided that the outer membrane is impermeable to Ca 2+ efflux into the cytoplasm. In this study, we demonstrate for the first time that the nuclear-encoded NCX3 isoform (1) is located on the outer mitochondrial membrane (OMM) of neurons; (2) colocalizes and immunoprecipitates with AKAP121 (also known as AKAP1), a member of the protein kinase A anchoring proteins (AKAPs) present on the outer membrane; (3) extrudes Ca 2+ from mitochondria through AKAP121 interaction in a PKA-mediated manner, both under normoxia and hypoxia; and (4) improves cell survival when it works in the Ca 2+ efflux mode at the level of the OMM. Collectively, these results suggest that, in neurons, NCX3 regulates mitochondrial Ca 2+ handling from the OMM through an AKAP121-anchored signaling complex, thus promoting cell survival during hypoxia.

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“…In the SOD2 MTS ATP6-39UTR SOD2 vector, both the MTS and the 39UTR of SOD2 were associated with the ATP6 ORF. The 39UTR of 215 bp (Ginsberg et al 2003) was inserted at the end of the ORF following the Flag epitope and replacing the SV40 poly(A) signal.…”

Section: Mtsmentioning

confidence: 99%

“…We have previously shown that SOD2 mRNA localizes to the mitochondrial surface in human cells (Sylvestre et al 2003a). Furthermore, a recent report described that in HeLa cells, SOD2 mRNA is associated to the mitochondrial surface, via its 39UTR linked to the K homology domain of A-kinase anchor protein 121, AKAP121 (Ginsberg et al 2003). HeLa cells were stably transfected with two different plasmids.…”

Section: Introductionmentioning

confidence: 99%

mRNA localization to the mitochondrial surface allows the efficient translocation inside the organelle of a nuclear recoded ATP6 protein

Kaltimbacher

Bonnet²,

Lecoeuvre³

et al. 2006

RNA

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As previously established in yeast, two sequences within mRNAs are responsible for their specific localization to the mitochondrial surface-the region coding for the mitochondrial targeting sequence and the 39UTR. This phenomenon is conserved in human cells. Therefore, we decided to use mRNA localization as a tool to address to mitochondria, a protein that is not normally imported. For this purpose, we associated a nuclear recoded ATP6 gene with the mitochondrial targeting sequence and the 39UTR of the nuclear SOD2 gene, which mRNA exclusively localizes to the mitochondrial surface in HeLa cells. The ATP6 gene is naturally located into the organelle and encodes a highly hydrophobic protein of the respiratory chain complex V.In this study, we demonstrated that hybrid ATP6 mRNAs, as the endogenous SOD2 mRNA, localize to the mitochondrial surface in human cells. Remarkably, fusion proteins localize to mitochondria in vivo. Indeed, ATP6 precursors synthesized in the cytoplasm were imported into mitochondria in a highly efficient way, especially when both the MTS and the 39UTR of the SOD2 gene were associated with the re-engineered ATP6 gene. Hence, these data indicate that mRNA targeting to the mitochondrial surface represents an attractive strategy for allowing the mitochondrial import of proteins originally encoded by the mitochondrial genome without any amino acid change in the protein that could interfere with its biologic activity.

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PKA-dependent Binding of mRNA to the Mitochondrial AKAP121 Protein

Cited by 94 publications

References 41 publications

Differential targeting of VDAC3 mRNA isoforms influences mitochondria morphology

Differential targeting of VDAC3 mRNA isoforms influences mitochondria morphology

NCX3 regulates mitochondrial calcium handling through AKAP121-anchored signaling complex and prevents hypoxia-induced cell death

mRNA localization to the mitochondrial surface allows the efficient translocation inside the organelle of a nuclear recoded ATP6 protein

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