Yeast mrna cap- binding protein Cbc1/Sto1 is necessary for the rapid reprogramming of transla9on a:er hyperosmo9c shock (2012)
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1 Transla'on in Eucaryotes Yeast mrna cap- binding protein Cbc1/Sto1 is necessary for the rapid reprogramming of transla9on a:er hyperosmo9c shock (2012) Elena Garrea, Lorena Romero- Santacreua, Nikki De Clercqa, Na9 Blasco- Anguloa, Per Sunnerhagenb, and Paula Alepuza University of València/ Goethenburg Maurus Beeli University of Bern, 2012
2 Introduc'on Stress signals - > shutdown and reprogramming of gene expression Protec9on of cell, adap9on to new environment mrnas for stress protec9ve proteins are selec9vely encoded Defects lead to diseases like cancer, diabetes and inflammatory processes (Keene, 2007) Mechanism and factors involved in the specific transla9on of osmostress- responsive genes in S. Cerevisiae unknown
3 Introduc'on Stress usually inhibits expression at ini9a9on step cap- dependent ini9a9on mechanism elf4e recognizes and binds to the cap structure of mrna
4 Introduc'on
5 Introduc'on Inhibitory proteins can bind elf4e to impede mrna binding mrnas encoding stress proteins have to be be translated Recruitment of ribosomes in 5 and 3 UTR s of mrnas, including IRES Upstream open reading frames MicroRNA target sites
6 Introduc'on Model: Hyperosmo9c shock in yeast Rapid decrease in protein synthesis HOG mitogenac9vated protein kinase (MAPK) pathway HOG1 ac9vated by phosphoryla9on 600 mrna levels change within minutes (10% of genes)
7 Introduc'on Nuclear Cap binding complex (CBC) Heterodimer, Cbc1/Cbc2 CBP80 in mammals Binds to cap structure of mrnas in the nucleus Mainly located in the nucleus, can shuele to the cytoplasm, using the impor9n α/β system Involved in splicing, transcrip9onal termina9on, protec9on of mrna degrada9on, role in transla9on?
8 Experimental procedure Experiment 1 Involved proteins Iden9fy proteins, involved in poseranscrip9onal regula9on under osmo9c stress Osmo9c stress sensi9vity screening 182 mutants in yeast genes A dele9on in Cbc1 was very sensi9ve, sugges9ng that Cbc1 is involved stress managing
9 Experiment 1 Cbc1 mutants showed a strong reduc9on in viability
10 Experiment 2 The P- bodies Change in transla9on level upon osmo9c shock Protein synthesis restored through unknown Hog1 mechanism Transient increase in P- bodies (lasts longer in Hog1 mutants) Storage of mrna un9l stress phase is over
11 Experiment 2 P- bodies were build in all strains Cbc1 not essen9al for P- body forma9on P- bodies last longer in Cbc1 mutants Cbc1 plays a role in reini9a9on of transla9on
12 Experiment 3 elf4e vs. Cbc1 elf4e replaces Cbc1 in cap structure of cytoplasmic mrnas elf4e is essen9al for cell growth Cbc1 under op9mal condi9ons not essen9al Cbc1 dele9on and thermosensi9ve muta9on in elf4e
13 Experiment 3 elf4e vs. Cbc1 Cdc33 ts = thermosensi've muta'on in elf4e Under osmo9c stress, Cbc1 takes the role of elf4e
14 Further inves'ga'ons Difference between Cbc1- mediated transla9on and elf4e- mediated transla9on Is Cbc1- dependent transla9on restricted to specific transcripts such as osmostress- responsive mrna? Is Cbc1 also involved in trasla9on reprogramming under other stress condi9ons?
15 Take Home messages In response to osmo9c stress, global transla9on is inhibited in yeast Cbc1, an mrna binding protein, is essen9al in this process Cbc1 is not essen9al under normal condi9ons and can be replaced by elf4e in the cytoplasm Cbc1- mediated transla9on is stress resistant while elf4e- mediated transla9on is stress- sensi9ve
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