Journal
NUCLEIC ACIDS RESEARCH
Volume 43, Issue 16, Pages 8013-8032Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkv731
Keywords
-
Categories
Funding
- German Research Foundation DFG [Kr4017/1-1, Kr4017/1-2, SI 1610/2-1]
- Young Investigator Program of the Research Center of Infectious Diseases (ZINF) of the University of Wurzburg, the Germany
- Human Frontier Science Program (TNS)
- Wellcome Trust [085956/Z/08/Z]
- DFG (German research association)
Ask authors/readers for more resources
RNP granules are ribonucleoprotein assemblies that regulate the post-transcriptional fate of mRNAs in all eukaryotes. Their exact function remains poorly understood, one reason for this is that RNP granule purification has not yet been achieved. We have exploited a unique feature of trypanosomes to prepare a cellular fraction highly enriched in starvation stress granules. First, granules remain trapped within the cage-like, subpellicular microtubule array of the trypanosome cytoskeleton while soluble proteins are washed away. Second, the microtubules are depolymerized and the granules are released. RNA sequencing combined with single molecule mRNA FISH identified the short and highly abundant mRNAs encoding ribosomal mRNAs as being excluded from granules. By mass spectrometry we have identified 463 stress granule candidate proteins. For 17/49 proteins tested by eYFP tagging we have confirmed the localization to granules, including one phosphatase, one methyltransferase and two proteins with a function in trypanosome life-cycle regulation. The novel method presented here enables the unbiased identification of novel RNP granule components, paving the way towards an understanding of RNP granule function.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available