Document Detail

In Vitro Reconstitution of a Cellular Phase-Transition Process that Involves the mRNA Decapping Machinery.
MedLine Citation:
PMID:  24862735     Owner:  NLM     Status:  Publisher    
In eukaryotic cells, components of the 5' to 3' mRNA degradation machinery can undergo a rapid phase transition. The resulting cytoplasmic foci are referred to as processing bodies (P-bodies). The molecular details of the self-aggregation process are, however, largely undetermined. Herein, we use a bottom-up approach that combines NMR spectroscopy, isothermal titration calorimetry, X-ray crystallography, and fluorescence microscopy to probe if mRNA degradation factors can undergo phase transitions in vitro. We show that the Schizosaccharomyces pombe Dcp2 mRNA decapping enzyme, its prime activator Dcp1, and the scaffolding proteins Edc3 and Pdc1 are sufficient to reconstitute a phase-separation process. Intermolecular interactions between the Edc3 LSm domain and at least 10 helical leucine-rich motifs in Dcp2 and Pdc1 build the core of the interaction network. We show that blocking of these interactions interferes with the clustering behavior, both in vitro and in vivo.
Simon A Fromm; Julia Kamenz; Erik R Nöldeke; Ancilla Neu; Georg Zocher; Remco Sprangers
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-5-26
Journal Detail:
Title:  Angewandte Chemie (International ed. in English)     Volume:  -     ISSN:  1521-3773     ISO Abbreviation:  Angew. Chem. Int. Ed. Engl.     Publication Date:  2014 May 
Date Detail:
Created Date:  2014-5-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370543     Medline TA:  Angew Chem Int Ed Engl     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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