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Bogart, B. M.

Publications and source records attributed to Bogart, B. M..

2 recordsLinked to original sources

Molecular Basis for Impacts of DSIF on the Dynamics of RNA Polymerase II Elongation Complex

Transcription elongation is a highly regulated process involving elongation factors associated with RNA polymerase II (Pol II). DRB sensitivity-inducing factor (DSIF) is an elongation factor known to have multiple roles in transcription elongation. Although studies resolved the structures of elongation complexes with DSIF, little is known about the impacts of DSIF on the dynamics of the elongation complex at the molecular level. Here, we used molecular dynamics simulations to elucidate the effects of DSIF on the dynamics and structure of Pol II and upstream nucleic acids, thereby gaining a mechanistic understanding of its role in transcription elongation. We determined three major sites of impact by DSIF, including the upstream nucleic acids, Pol II clamp, and active site, which potentially contribute to its role in transcription processivity. Our results showed that DSIF affects the dynamics of upstream DNA and RNA at the exit sites, preventing the unwinding of DNA and the folding of RNA. In addition, our results suggest that DSIF regulates the motion of the Pol II clamp to potentially maintain a proper size at the central cleft. We also observed a more dynamic active site and increased interactions between active site domains. Based on correlated motion analysis, we proposed that the impacts of DSIF on the active site have an allosteric nature that takes place through the collective motions of the Pol II clamp and nucleic acids.

biophysics↗

Protein-Nucleic Acid Interactions for RNA Polymerase II Elongation Factors by Molecular Dynamics Simulations

RNA polymerase II (Pol II) forms a complex with elongation factors to proceed the elongation stage of the transcription process. In this work, we studied elongation factor SPT5 and explored protein nucleic acid interactions for the isolated systems of KOW1 and KOW4 domains of SPT5 with DNA and RNA, respectively. We performed molecular dynamics (MD) simulations using three commonly used force fields that are CHARMM c36m, AMBER ff14sb and ff19sb. These simulations showed that most of the protein-nucleic acid interactions in the native state were retained with an increased electrostatic binding free energy for all force fields used. RNA was found highly dynamic with all force fields while DNA had relatively more stable conformations with the AMBER force fields compared to CHARMM. Furthermore, we performed MD simulations of the complete elongation complex using CHARMM c36m force field to compare the dynamics and interactions in the isolated systems. Similar strong KOW1 and DNA interactions were observed in the complete elongation complex simulations and DNA was further stabilized by a network of interactions involving SPT5-KOW1, SPT4 and rpb2 of Pol II. Overall, our study showed that the accuracy of force fields and the presence of the entire interaction network are important for elucidating the dynamics of protein-nucleic acid systems.

biophysics↗