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The DEAD-box helicase Ded1 from yeast is an mRNP cap-associated protein that shuttles between the cytoplasm and nucleus.

Senissar M, Le Saux A, Belgareh-Touzé N, Adam C, Banroques J, Tanner NK - Nucleic Acids Res. (2014)

Bottom Line: In addition, we show that Ded1 is genetically linked to these factors.Ded1 comigrates with these proteins on sucrose gradients, but treatment with rapamycin does not appreciably alter the distribution of Ded1; thus, most of the Ded1 is in stable mRNP complexes.We conclude that Ded1 is an mRNP cofactor of the cap complex that may function to remodel the different mRNPs and thereby regulate the expression of the mRNAs.

View Article: PubMed Central - PubMed

Affiliation: Expression Génétique Microbienne, CNRS FRE3630 (UPR9073), in association with Université Paris Diderot, Sorbonne Paris Cité, Paris 75005, France Université Paris-Sud, Ecole Doctorale 426 GGC, Orsay, France.

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Protein partners of Ded1. Ded1 interacts with the predominately nuclear CBC complex factors, which consists of Cbp20 and Cbp80, and with Nab2. Ded1 also interacts with the predominately cytoplasmic translation-initiation factors, which includes Pab1. We have no evidence that Ded1 interacts with the DEAD-box protein eIF4A, which is part of the translation-initiation complex eIF4F. Moreover, Ded1 interacts with the DEAD-box protein Dhh1, which is often localized in P-bodies in the cytoplasm. It is involved in mRNA decapping and degradation. Gle1 is located on the cytoplasmic side of the nuclear pore, and it is involved in the nuclear export of mRNAs. However, Gle1 also has been implicated in regulating translation. The heavy lines represent interactions determined by physical, genetic and enzymatic approaches, the medium line by physical and genetic approaches, and the medium dotted line by physical and enzymatic approaches. The light dotted line indicates other potential substrates. Interactions were determined by this work and the published work of others (see text).
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Figure 7: Protein partners of Ded1. Ded1 interacts with the predominately nuclear CBC complex factors, which consists of Cbp20 and Cbp80, and with Nab2. Ded1 also interacts with the predominately cytoplasmic translation-initiation factors, which includes Pab1. We have no evidence that Ded1 interacts with the DEAD-box protein eIF4A, which is part of the translation-initiation complex eIF4F. Moreover, Ded1 interacts with the DEAD-box protein Dhh1, which is often localized in P-bodies in the cytoplasm. It is involved in mRNA decapping and degradation. Gle1 is located on the cytoplasmic side of the nuclear pore, and it is involved in the nuclear export of mRNAs. However, Gle1 also has been implicated in regulating translation. The heavy lines represent interactions determined by physical, genetic and enzymatic approaches, the medium line by physical and genetic approaches, and the medium dotted line by physical and enzymatic approaches. The light dotted line indicates other potential substrates. Interactions were determined by this work and the published work of others (see text).

Mentions: Our experiments demonstrate that Ded1 is a physical and functional component of the nuclear and cytoplasmic cap-binding complexes. We find that Ded1 is genetically linked to various CBC and eIF4F proteins, and that it physically interacts with these proteins in vitro (Figure 7). Moreover, we find that these proteins enhance the RNA-dependent ATPase activity of Ded1 in vitro. The Ded1-GFP protein actively shuttles between the nucleus and cytoplasm using both the Crm1- and Mex67-dependent nuclear export pathways, which is largely independent of the enzymatic activity of Ded1. Thus, Ded1 is probably a component of RNPs located both in the nucleus and the cytoplasm. Our mass spectrometry analyses indicate that Ded1 is associated with a large number of other proteins in cell extracts and thus in multiple different RNPs. Finally, sucrose gradients reveal that Ded1 cosediments with known cap-associated factors but that the peak concentrations of the factors do not coincide, which indicates that Ded1 interacts with a subset of these factors that form larger complexes. This distribution is not profoundly affected with rapamycin treatment, which indicates that the majority of the RNPs associated with Ded1 are not actively being translated.


The DEAD-box helicase Ded1 from yeast is an mRNP cap-associated protein that shuttles between the cytoplasm and nucleus.

Senissar M, Le Saux A, Belgareh-Touzé N, Adam C, Banroques J, Tanner NK - Nucleic Acids Res. (2014)

Protein partners of Ded1. Ded1 interacts with the predominately nuclear CBC complex factors, which consists of Cbp20 and Cbp80, and with Nab2. Ded1 also interacts with the predominately cytoplasmic translation-initiation factors, which includes Pab1. We have no evidence that Ded1 interacts with the DEAD-box protein eIF4A, which is part of the translation-initiation complex eIF4F. Moreover, Ded1 interacts with the DEAD-box protein Dhh1, which is often localized in P-bodies in the cytoplasm. It is involved in mRNA decapping and degradation. Gle1 is located on the cytoplasmic side of the nuclear pore, and it is involved in the nuclear export of mRNAs. However, Gle1 also has been implicated in regulating translation. The heavy lines represent interactions determined by physical, genetic and enzymatic approaches, the medium line by physical and genetic approaches, and the medium dotted line by physical and enzymatic approaches. The light dotted line indicates other potential substrates. Interactions were determined by this work and the published work of others (see text).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 7: Protein partners of Ded1. Ded1 interacts with the predominately nuclear CBC complex factors, which consists of Cbp20 and Cbp80, and with Nab2. Ded1 also interacts with the predominately cytoplasmic translation-initiation factors, which includes Pab1. We have no evidence that Ded1 interacts with the DEAD-box protein eIF4A, which is part of the translation-initiation complex eIF4F. Moreover, Ded1 interacts with the DEAD-box protein Dhh1, which is often localized in P-bodies in the cytoplasm. It is involved in mRNA decapping and degradation. Gle1 is located on the cytoplasmic side of the nuclear pore, and it is involved in the nuclear export of mRNAs. However, Gle1 also has been implicated in regulating translation. The heavy lines represent interactions determined by physical, genetic and enzymatic approaches, the medium line by physical and genetic approaches, and the medium dotted line by physical and enzymatic approaches. The light dotted line indicates other potential substrates. Interactions were determined by this work and the published work of others (see text).
Mentions: Our experiments demonstrate that Ded1 is a physical and functional component of the nuclear and cytoplasmic cap-binding complexes. We find that Ded1 is genetically linked to various CBC and eIF4F proteins, and that it physically interacts with these proteins in vitro (Figure 7). Moreover, we find that these proteins enhance the RNA-dependent ATPase activity of Ded1 in vitro. The Ded1-GFP protein actively shuttles between the nucleus and cytoplasm using both the Crm1- and Mex67-dependent nuclear export pathways, which is largely independent of the enzymatic activity of Ded1. Thus, Ded1 is probably a component of RNPs located both in the nucleus and the cytoplasm. Our mass spectrometry analyses indicate that Ded1 is associated with a large number of other proteins in cell extracts and thus in multiple different RNPs. Finally, sucrose gradients reveal that Ded1 cosediments with known cap-associated factors but that the peak concentrations of the factors do not coincide, which indicates that Ded1 interacts with a subset of these factors that form larger complexes. This distribution is not profoundly affected with rapamycin treatment, which indicates that the majority of the RNPs associated with Ded1 are not actively being translated.

Bottom Line: In addition, we show that Ded1 is genetically linked to these factors.Ded1 comigrates with these proteins on sucrose gradients, but treatment with rapamycin does not appreciably alter the distribution of Ded1; thus, most of the Ded1 is in stable mRNP complexes.We conclude that Ded1 is an mRNP cofactor of the cap complex that may function to remodel the different mRNPs and thereby regulate the expression of the mRNAs.

View Article: PubMed Central - PubMed

Affiliation: Expression Génétique Microbienne, CNRS FRE3630 (UPR9073), in association with Université Paris Diderot, Sorbonne Paris Cité, Paris 75005, France Université Paris-Sud, Ecole Doctorale 426 GGC, Orsay, France.

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