Novel insights into RNP granules by employing the trypanosome's microtubule skeleton as a molecular sieve.

摘要: 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 granule purification has not yet been achieved. We have exploited a unique feature trypanosomes to prepare cellular fraction highly enriched starvation stress granules. First, remain trapped within cage-like, subpellicular microtubule array trypanosome cytoskeleton while soluble proteins washed away. Second, microtubules depolymerized and released. RNA sequencing combined with single molecule mRNA FISH identified short abundant encoding ribosomal as being excluded from By mass spectrometry we 463 candidate proteins. For 17/49 tested by eYFP tagging confirmed localization granules, including phosphatase, methyltransferase two life-cycle regulation. The novel method presented here enables unbiased identification components, paving way towards an understanding function.