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Scientific article
Open access
English

The Not5 Subunit of the Ccr4-Not Complex Connects Transcription and Translation

Published inPLOS genetics, vol. 10, no. 10, e1004569
Publication date2014
Abstract

Recent studies have suggested that a sub-complex of RNA polymerase II composed of Rpb4 and Rpb7 couples the nuclear and cytoplasmic stages of gene expression by associating with newly made mRNAs in the nucleus, and contributing to their translation and degradation in the cytoplasm. Here we show by yeast two hybrid and co-immunoprecipitation experiments, followed by ribosome fractionation and fluorescent microscopy, that a subunit of the Ccr4-Not complex, Not5, is essential in the nucleus for the cytoplasmic functions of Rpb4. Not5 interacts with Rpb4; it is required for the presence of Rpb4 in polysomes, for interaction of Rpb4 with the translation initiation factor eIF3 and for association of Rpb4 with mRNAs. We find that Rpb7 presence in the cytoplasm and polysomes is much less significant than that of Rpb4, and that it does not depend upon Not5. Hence Not5-dependence unlinks the cytoplasmic functions of Rpb4 and Rpb7. We additionally determine with RNA immunoprecipitation and native gel analysis that Not5 is needed in the cytoplasm for the co-translational assembly of RNA polymerase II. This stems from the importance of Not5 for the association of the R2TP Hsp90 co-chaperone with polysomes translating RPB1 mRNA to protect newly synthesized Rpb1 from aggregation. Hence taken together our results show that Not5 interconnects translation and transcription.

Keywords
  • Ccr4-Not complex
  • Not5
  • Rpb4
  • RNA polymerase II
  • Complex assembly
  • Yeast
  • Transcription
  • Translation
  • R2TP
  • Hsp90
  • Chaperone
Citation (ISO format)
VILLANYI, Zoltan et al. The Not5 Subunit of the Ccr4-Not Complex Connects Transcription and Translation. In: PLOS genetics, 2014, vol. 10, n° 10, p. e1004569. doi: 10.1371/journal.pgen.1004569
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Article (Published version)
accessLevelPublic
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ISSN of the journal1553-7390
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