In vivo SELEX reveals novel sequence and structural determinants of Nrd1-Nab3-Sen1 dependent transcription termination

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Authors

PORRUA-FUERTE Odil HÓBOR Fruzsina BOULAY Jocelyne KUBÍČEK Karel D’AUBENTON-CARAFA Yves GUDIPATI Rajani Kanth ŠTEFL Richard LIBRI Domenico

Year of publication 2012
Type Article in Periodical
Magazine / Source EMBO Journal
MU Faculty or unit

Central European Institute of Technology

Citation
Web http://www.nature.com/emboj/journal/v31/n19/abs/emboj2012237a.html
Doi http://dx.doi.org/10.1038/emboj.2012.237
Field Genetics and molecular biology
Keywords cryptic unstable transcripts; transcription termination; Nrd1p-Nab3p-Sen1p complex; hidden transcription; transcriptional insulators
Description The Nrd1-Nab3-Sen1 (NNS) complex pathway is responsible for transcription termination of cryptic unstable transcripts and sn/snoRNAs. The NNS complex recognizes short motifs on the nascent RNA, but the presence of these sequences alone is not sufficient to define a functional terminator. We generated a homogeneous set of several hundreds of artificial, NNS-dependent terminators with an in vivo selection approach. Analysis of these terminators revealed novel and extended sequence determinants for transcription termination and NNS complex binding as well as supermotifs that are critical for termination. Biochemical and structural data revealed that affinity and specificity of RNA recognition by Nab3p relies on induced fit recognition implicating an a-helical extension of the RNA recognition motif. Interestingly, the same motifs can be recognized by the NNS or the mRNA termination complex depending on their position relative to the start of transcription, suggesting that they function as general transcriptional insulators to prevent interference between the non-coding and the coding yeast transcriptomes.
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