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Xenopus laevis as a Model to Identify Translation Impairment.
de Broucker A
,
Semaille P
,
Cailliau K
,
Martoriati A
,
Comptdaer T
,
Bodart JF
,
Destée A
,
Chartier-Harlin MC
.
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Protein synthesis is a fundamental process to gene expression impacting diverse biological processes notably adaptation to environmental conditions. The initiation step, which involves the assembly of the ribosomal subunits at the mRNA initiation codon, involved initiation factor including eIF4G1. Defects in this rate limiting step of translation are linked to diverse disorders. To study the potential consequences of such deregulations, Xenopus laevis oocytes constitute an attractive model with high degrees of conservation of essential cellular and molecular mechanisms with human. In addition, during meiotic maturation, oocytes are transcriptionally repressed and all necessary proteins are translated from preexisting, maternally derived mRNAs. This inexpensive model enables exogenous mRNA to become perfectly integrated with an effective translation. Here is described a protocol for assessing translation with a factor of interest (here eIF4G1) using stored maternal mRNA that are the first to be polyadenylated and translated during oocyte maturation as a physiological readout. At first, mRNA synthetized by in vitro transcription of plasmids of interest (here eIF4G1) are injected in oocytes and kinetics of oocyte maturation by Germinal Vesicle Breakdown detection is determined. The studied maternal mRNA target is the serine/threonine-protein-kinase mos. Its polyadenylation and its subsequent translation are investigated together with the expression and phosphorylation of proteins of the mos signaling cascade involved in oocyte maturation. Variations of the current protocol to put forward translational defects are also proposed to emphasize its general applicability. In light of emerging evidence that aberrant protein synthesis may be involved in the pathogenesis of neurological disorders, such a model provides the opportunity to easily assess this impairment and identify new targets.
Au,
Microinjection of Xenopus laevis oocytes as a system for studying nuclear transport of viruses.
2010, Pubmed,
Xenbase
Au,
Microinjection of Xenopus laevis oocytes as a system for studying nuclear transport of viruses.
2010,
Pubmed
,
Xenbase
Charlesworth,
Cytoplasmic polyadenylation element (CPE)- and CPE-binding protein (CPEB)-independent mechanisms regulate early class maternal mRNA translational activation in Xenopus oocytes.
2004,
Pubmed
,
Xenbase
Chartier-Harlin,
Translation initiator EIF4G1 mutations in familial Parkinson disease.
2011,
Pubmed
Cohen,
Microinjection of Xenopus laevis oocytes.
2009,
Pubmed
,
Xenbase
Doyle,
Unfolded proteins and endoplasmic reticulum stress in neurodegenerative disorders.
2011,
Pubmed
Dumont,
Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals.
1972,
Pubmed
,
Xenbase
Gandin,
Polysome fractionation and analysis of mammalian translatomes on a genome-wide scale.
2014,
Pubmed
Gebauer,
Synthesis and function of Mos: the control switch of vertebrate oocyte meiosis.
1997,
Pubmed
,
Xenbase
Gotoh,
Regulatory pathways coordinating cell cycle progression in early Xenopus development.
2011,
Pubmed
,
Xenbase
Holcik,
Translational control in stress and apoptosis.
2005,
Pubmed
Jackson,
The mechanism of eukaryotic translation initiation and principles of its regulation.
2010,
Pubmed
Keiper,
Translational recruitment of Xenopus maternal mRNAs in response to poly(A) elongation requires initiation factor eIF4G-1.
1999,
Pubmed
,
Xenbase
Lin,
Loss of PINK1 attenuates HIF-1α induction by preventing 4E-BP1-dependent switch in protein translation under hypoxia.
2014,
Pubmed
López-Lastra,
Protein synthesis in eukaryotes: the growing biological relevance of cap-independent translation initiation.
2005,
Pubmed
Martin,
Ribosomal protein s15 phosphorylation mediates LRRK2 neurodegeneration in Parkinson's disease.
2014,
Pubmed
Moreno,
Oral treatment targeting the unfolded protein response prevents neurodegeneration and clinical disease in prion-infected mice.
2013,
Pubmed
Méreau,
Xenopus as a model to study alternative splicing in vivo.
2007,
Pubmed
,
Xenbase
Rassa,
Spacing constraints on reinitiation of paramyxovirus transcription: the gene end U tract acts as a spacer to separate gene end from gene start sites.
2000,
Pubmed
Schmidt,
SUnSET, a nonradioactive method to monitor protein synthesis.
2009,
Pubmed
Senée,
Wolcott-Rallison Syndrome: clinical, genetic, and functional study of EIF2AK3 mutations and suggestion of genetic heterogeneity.
2004,
Pubmed
Sheets,
Polyadenylation of c-mos mRNA as a control point in Xenopus meiotic maturation.
1995,
Pubmed
,
Xenbase
Sonenberg,
Regulation of translation initiation in eukaryotes: mechanisms and biological targets.
2009,
Pubmed
Terenin,
A novel mechanism of eukaryotic translation initiation that is neither m7G-cap-, nor IRES-dependent.
2013,
Pubmed
Wakiyama,
Interaction of eIF4G with poly(A)-binding protein stimulates translation and is critical for Xenopus oocyte maturation.
2000,
Pubmed
,
Xenbase
Willis,
Study of the DNA damage checkpoint using Xenopus egg extracts.
2012,
Pubmed
,
Xenbase
van der Knaap,
Mutations in each of the five subunits of translation initiation factor eIF2B can cause leukoencephalopathy with vanishing white matter.
2002,
Pubmed
