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Title: Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS

Abstract

RNA-binding protein with multiple splicing (designated RBPMS) is a higher vertebrate mRNA-binding protein containing a single RNA recognition motif (RRM). RBPMS has been shown to be involved in mRNA transport, localization and stability, with key roles in axon guidance, smooth muscle plasticity, as well as regulation of cancer cell proliferation and migration. We report on structure-function studies of the RRM domain of RBPMS bound to a CAC-containing single-stranded RNA. These results provide insights into potential topologies of complexes formed by the RBPMS RRM domain and the tandem CAC repeat binding sites as detected by photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation. These studies establish that the RRM domain of RBPMS forms a symmetrical dimer in the free state, with each monomer binding sequence-specifically to all three nucleotides of a CAC segment in the RNA bound state. Structure-guided mutations within the dimerization and RNA-binding interfaces of RBPMS RRM on RNA complex formation resulted in both disruption of dimerization and a decrease in RNA-binding affinity as observed by size exclusion chromatography and isothermal titration calorimetry. Here, as anticipated from biochemical binding studies, over-expression of dimerization or RNA-binding mutants of Flag-HA-tagged RBPMS were no longer able to track with stress granules in HEK293 cells, thereby documentingmore » the deleterious effects of such mutations in vivo.« less

Authors:
 [1];  [2];  [2];  [1]
  1. Memorial Sloan-Kettering Cancer Center, New York, NY (United States)
  2. The Rockefeller Univ., New York, NY (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH); RU CCTS
OSTI Identifier:
1430282
Grant/Contract Number:  
R01GM104962; UL1RR024143
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Quarterly Reviews of Biophysics
Additional Journal Information:
Journal Volume: 49; Journal Issue: 2016; Journal ID: ISSN 0033-5835
Publisher:
Cambridge University Press
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; The RNA-binding protein with multiple splicing; RNA recognition motif; photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation; RNA recognition element

Citation Formats

Teplova, Marianna, Farazi, Thalia A., Tuschl, Thomas, and Patel, Dinshaw J. Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS. United States: N. p., 2015. Web. doi:10.1017/S0033583515000207.
Teplova, Marianna, Farazi, Thalia A., Tuschl, Thomas, & Patel, Dinshaw J. Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS. United States. doi:10.1017/S0033583515000207.
Teplova, Marianna, Farazi, Thalia A., Tuschl, Thomas, and Patel, Dinshaw J. Tue . "Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS". United States. doi:10.1017/S0033583515000207. https://www.osti.gov/servlets/purl/1430282.
@article{osti_1430282,
title = {Structural basis underlying CAC RNA recognition by the RRM domain of dimeric RNA-binding protein RBPMS},
author = {Teplova, Marianna and Farazi, Thalia A. and Tuschl, Thomas and Patel, Dinshaw J.},
abstractNote = {RNA-binding protein with multiple splicing (designated RBPMS) is a higher vertebrate mRNA-binding protein containing a single RNA recognition motif (RRM). RBPMS has been shown to be involved in mRNA transport, localization and stability, with key roles in axon guidance, smooth muscle plasticity, as well as regulation of cancer cell proliferation and migration. We report on structure-function studies of the RRM domain of RBPMS bound to a CAC-containing single-stranded RNA. These results provide insights into potential topologies of complexes formed by the RBPMS RRM domain and the tandem CAC repeat binding sites as detected by photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation. These studies establish that the RRM domain of RBPMS forms a symmetrical dimer in the free state, with each monomer binding sequence-specifically to all three nucleotides of a CAC segment in the RNA bound state. Structure-guided mutations within the dimerization and RNA-binding interfaces of RBPMS RRM on RNA complex formation resulted in both disruption of dimerization and a decrease in RNA-binding affinity as observed by size exclusion chromatography and isothermal titration calorimetry. Here, as anticipated from biochemical binding studies, over-expression of dimerization or RNA-binding mutants of Flag-HA-tagged RBPMS were no longer able to track with stress granules in HEK293 cells, thereby documenting the deleterious effects of such mutations in vivo.},
doi = {10.1017/S0033583515000207},
journal = {Quarterly Reviews of Biophysics},
issn = {0033-5835},
number = 2016,
volume = 49,
place = {United States},
year = {2015},
month = {9}
}

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Cited by: 9 works
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    Works referencing / citing this record:

    Identification of RBPMS as a mammalian smooth muscle master splicing regulator via proximity of its gene with super-enhancers
    journal, July 2019

    • Nakagaki-Silva, Erick E.; Gooding, Clare; Llorian, Miriam
    • eLife, Vol. 8
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