التفاصيل البيبلوغرافية
| العنوان: |
Context-dependence of T-loop Mediated Long-range RNA Tertiary Interactions. |
| المؤلفون: |
Hansen, Lisa N.1 (AUTHOR), Kletzien, Otto A.1 (AUTHOR), Urquijo, Marcus1 (AUTHOR), Schwanz, Logan T.1 (AUTHOR), Batey, Robert T.1 (AUTHOR) Robert.Batey@colorado.edu |
| المصدر: |
Journal of Molecular Biology. May2023, Vol. 435 Issue 10, pN.PAG-N.PAG. 1p. |
| مصطلحات موضوعية: |
*RNA, *FLAVIN mononucleotide, *LIGAND binding (Biochemistry), *SEQUENCE spaces, *REPORTER genes, *TRANSFER RNA |
| مستخلص: |
[Display omitted] • The T-loop motif frequently mediates tertiary interactions in RNA. • Activity-based screen used to explore T-loop mediated interactions in riboswitches. • Results from the screen were consistent with phylogenetic analysis. • Consensus sequence for three different T-loop receptor motifs. • Results show T-loop/T-loop receptor interactions are context dependent. The architecture and folding of complex RNAs is governed by a limited set of highly recurrent structural motifs that form long-range tertiary interactions. One of these motifs is the T-loop, which was first identified in tRNA but is broadly distributed across biological RNAs. While the T-loop has been examined in detail in different biological contexts, the various receptors that it interacts with are not as well defined. In this study, we use a cell-based genetic screen in concert with bioinformatic analysis to examine three different, but related, T-loop receptor motifs found in the flavin mononucleotide (FMN) and cobalamin (Cbl) riboswitches. As a host for different T-loop receptors, we employed the env 8 class-II Cbl riboswitch, an RNA that uses two T-loop motifs for both folding and supporting the ligand binding pocket. A set of libraries was created in which select nucleotides that participate in the T-loop/T-loop receptor (TL/TLR) interaction were fully randomized. Library members were screened for their ability to support Cbl-dependent expression of a reporter gene. While T-loops appear to be variable in sequence, we find that the functional sequence space is more restricted in the Cbl riboswitch, suggesting that TL/TLR interactions are context dependent. Our data reveal clear sequence signatures for the different types of receptor motifs that align with phylogenic analysis of these motifs in the FMN and Cbl riboswitches. Finally, our data suggest the functional contribution of various nucleobase-mediated long-range interactions within the riboswitch subclass of TL/TLR interactions that are distinct from those found in other RNAs. [ABSTRACT FROM AUTHOR] |
| قاعدة البيانات: |
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