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Project #6701
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#6701 : Identification of Tac4 mRNA targets
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Name of Applicant : Micheline Fromont-Racine
Date of application : 27-06-2016
Unit : Genetics of Macromolecular Interactions
Location : Fernbach
Phone : 0140613432
@ Mail : mfromont@pasteur.fr
@ PI-Mail : jacquier@pasteur.fr

Project context and summary :

We are interested in the cytoplasmic quality control of gene expression and more especially into the behavior of aberrant peptides which could be generated from non-conform translation events. We are now investigating the role of a Saccharomyces cerevisiae RNA helicase protein that we named Tac4 (for Translation associated Component 4). We showed that this protein is involved in translation. We demonstrated, by sucrose gradient and affinity purification that Tac4 interacts with the ribosome. A first UV cross-linking and cDNA analysis (CRAC) experiment clearly revealed that Tac4 interacts with the 18S rRNA of the 40S ribosomal subunit and we precisely defined the crosslink point. These preliminary results also suggested an enrichment of the 3’-end regions of mRNAs. This implies that Tac4 could not only interact with the small ribosomal subunit but also directly with mRNA. Tac4 is conserved through the evolution and its mammalian homologue is involved in initiation of translation. Therefore, we thought that Tac4 could be associated with the 5’-end rather than with the 3’-end. However, a recent paper from the Rachel Green’s lab showed that translation reinitiation into the 3’-UTR region may occurs when translation termination is affected (Young et al., Cell 2015). The factors and molecular mechanisms implicated in these events are not known. Altogether, our preliminary results suggest that Tac4 is an excellent candidate participating to the unwinding of RNA structure or to the release of some RNA-binding proteins into the 3’-end mRNA. We now would like to 1) confirm that Tac4 preferentially interacts with the 3’-end of mRNA, 2) determine whether Tac4 interacts with a region upstream the Stop codon or in the 3’-UTR of the mRNA, 3) identify the mRNA targets to determine whether Tac4 could have a general role in translation or could only be involved in translation of some specific mRNA.


Related team publications :
Defenouillère, Q., E. Zhang, A. Namane, J. Mouaikel, A. Jacquier and M. Fromont-Racine. 2016. Rqc1 and Ltn1 prevent C-terminal Alanine-Threonine tail (CAT-tail) induced protein aggregation by efficient recruitment of Cdc48 on stalled 60S subunits. J. B. C. 291(23):12245-53
Defenouillère, Q.*, Y. Yao*, J. Mouaikel, A. Namane, A. Galopier, L. Decourty, A. Doyen, C. Malabat, C. Saveanu, A. Jacquier*, and M. Fromont-Racine*. 2013. Cdc48-associated complex bound to 60S particles is required for the clearance of aberrant translation products. Proc Natl Acad Sci USA 110: 5046-5051.
Babiano, R.*, G. Badis *, C. Saveanu, A. Namane, A. Doyen, A. Díaz-Quintana, A. Jacquier, Fromont-Racine, M.*, and J. de la Cruz*. 2013. Yeast ribosomal protein L7 and its homologue Rlp7 are simultaneously present at distinct sites on pre-60S ribosomal particles. Nucleic Acids Res 41(20):9461-70.
Service Delivery
Project Manager : varun.khanna@pasteur.fr
Project Type : Short
Status : In Progress


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