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Masipeddi, S.

Publications and source records attributed to Masipeddi, S..

2 recordsLinked to original sources

Deciphering conformational preferences of RNA in protein-RNA recognition

Conformational plasticity of RNAs plays important roles in recognizing RNA-binding proteins, and is often modulated by their binding partners. Here, we investigate RNA conformational preferences in a non-redundant dataset of 263 protein-RNA complexes to characterize the structural landscape associated with protein recognition. RNA dinucleotide segments are analyzed using seven backbone torsion angles ({delta}1, {varepsilon}1, {zeta}1, 2, {beta}2, {gamma}2, and {delta}2), two glycosidic torsion angles ({chi}1 and {chi}2) and the pseudo-torsion angle . Focusing on dinucleotide steps present in both interface and non-interface regions, we performed density-based clustering using selected backbone torsion angles to identify recurrent conformational states. We identify 28 distinct RNA dinucleotide conformers containing at least ten members each. Among these, eight conformers represent previously unreported nucleotide conformers (NtCs), including the transitional and the non-canonical states AB06, AB07, BB21, BB22, OP32, OP33, IC08 and IC09. Several of these conformers are preferentially enriched at protein-binding interfaces, suggesting their involvement in local conformational adaptation during protein-RNA recognition. The newly identified conformers span transitional A-B geometries, distorted B-like states, open conformations and compact intercalated structures, highlighting the remarkable structural plasticity of RNA in ribonucleoprotein complexes. Overall, this study expands the current understanding of RNA conformational space and provides a refined RNA dinucleotide conformer library for protein-RNA complexes. These findings will facilitate the identification of novel RNA structural motifs and improved RNA structural modeling, docking protein-RNA complexes and deep learning-based prediction frameworks for describing RNA tertiary structures.

biophysics↗

Decoding Mutually Induced Conformational Changes in Non-Canonical Recognition of U1 SL4 snRNA by ULD of SF3A1 during Early Spliceosome Assembly

A crucial step in early spliceosome assembly is the interaction between splicing factor 3A1 (SF3A1) of U2 snRNP and stem-loop 4 (SL4) of U1 snRNA. This interaction facilitates the spatial alignment of the 5' and the 3' splice sites, leading to the formation of the pre-spliceosomal A complex. In this study, we investigate the structural and dynamic basis of non-canonical recognition between ubiquitin-like domain (ULD) of SF3A1 and SL4 of U1 snRNA. Extensive all-atom molecular dynamics simulations reveal a dual recognition mechanism involving sequence-specific interactions mediated by the C-terminal RGGR motif and structural recognition governed by the UUCG tetraloop of SL4 snRNA. The RGGR motif primarily engages the duplex region of the snRNA, whereas the stem-loop nucleotides interact with the globular region of the ULD. Mutations of key residues R788 and R791 result in a significant loss of protein-RNA interactions, as reflected in the reduced binding affinities and altered conformational stability. Nucleotides C6 to C9 in the duplex region, stabilized by strong base-pairing and backbone-mediated interactions with SF3A1, exhibit constrained torsional distributions and minimal sensitivity to mutation. In contrast, nucleotides G10 to C15 in the tetraloop exhibit broader torsional distributions with moderate occupancy, consistent with weaker base-pairing but stronger protein-RNA interactions. Mutations significantly alter torsional distribution, enabling the nucleotides to adopt alternate conformations that preserve interactions with the globular domain of SF3A1. These findings provide a mechanistic insight into non-canonical RNA recognition and highlight the role of coupled sequence and structural determinants in stabilizing early spliceosomal assembly.

biophysics↗