西亚试剂：Single-molecule analysis of Mss116-mediated group II intron

发布时间：2026-01-28

Single-molecule analysis of Mss116-mediated group II intron folding
Krishanthi S. Karunatilaka1, Amanda Solem1, Anna Marie Pyle2,3 & David Rueda1

1Department of Chemistry, Wayne State University, 5101 Cass Avenue, Detroit, Michigan 48202, USA
2Department of Molecular, Cellular and Developmental Biology and Department of Chemistry, Yale University, New Haven, Connecticut 06520, USA
3Howard Hughes Medical Institute, Yale University, New Haven, Connecticut 06520, USA

DEAD-box helicases are conserved enzymes involved in nearly all aspects of RNA metabolism, but their mechanisms of action remain unclear. Here, we investigated the mechanism of the DEAD-box protein Mss116 on its natural substrate, the group II intron ai5γ. Group II introns are structurally complex catalytic RNAs considered evolutionarily related to the eukaryotic spliceosome, and an interesting paradigm for large RNA folding. We used single-molecule fluorescence to monitor the effect of Mss116 on folding dynamics of a minimal active construct, ai5γ?D135. The data show that Mss116 stimulates dynamic sampling between states along the folding pathway, an effect previously observed only with high Mg2+ concentrations. Furthermore, the data indicate that Mss116 promotes folding through discrete ATP-independent and ATP-dependent steps. We propose that Mss116 stimulates group II intron folding through a multi-step process that involves electrostatic stabilization of early intermediates and ATP hydrolysis during the final stages of native state assembly.

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