Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3)

Kashyap Maruthi, Ashwani Sharma, Neel Sarovar Bhavesh

Research output: Contribution to journalArticle

Abstract

Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P213, with unit-cell parameters a = b = c = 118.5 Å, α = β = γ = 90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (V M) was 6.35 Å3 Da -1, with a solvent content of 80.62%. Initial phases were determined by molecular replacement.

Original languageEnglish (US)
Pages (from-to)1107-1109
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume69
Issue number10
DOIs
StatePublished - Oct 1 2013
Externally publishedYes

Fingerprint

RNA-Binding Proteins
Crystallization
purification
Purification
RNA
crystallization
proteins
Troponin T
RNA Precursors
Alternative Splicing
editing
splicing
Exons
muscles
Muscles
inclusions
Muscle
vapors
Vapors
RNA Recognition Motif

Keywords

  • C-terminal RNA recognition motif
  • ETR-3
  • human ELAV-type RNA-binding protein 3
  • RRM-3

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

@article{b9b4abfdeb304cbca48dd20e061a0d5a,
title = "Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3)",
abstract = "Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 {\AA} resolution at the home source and belonged to space group P213, with unit-cell parameters a = b = c = 118.5 {\AA}, α = β = γ = 90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (V M) was 6.35 {\AA}3 Da -1, with a solvent content of 80.62{\%}. Initial phases were determined by molecular replacement.",
keywords = "C-terminal RNA recognition motif, ETR-3, human ELAV-type RNA-binding protein 3, RRM-3",
author = "Kashyap Maruthi and Ashwani Sharma and Bhavesh, {Neel Sarovar}",
year = "2013",
month = "10",
day = "1",
doi = "10.1107/S1744309113023439",
language = "English (US)",
volume = "69",
pages = "1107--1109",
journal = "Acta Crystallographica Section F:Structural Biology Communications",
issn = "1744-3091",
publisher = "John Wiley and Sons Ltd",
number = "10",

}

TY - JOUR

T1 - Purification, crystallization and preliminary crystallographic studies of C-terminal RNA recognition motif (RRM-3) of human ELAV-type RNA-binding protein 3 (ETR-3)

AU - Maruthi, Kashyap

AU - Sharma, Ashwani

AU - Bhavesh, Neel Sarovar

PY - 2013/10/1

Y1 - 2013/10/1

N2 - Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P213, with unit-cell parameters a = b = c = 118.5 Å, α = β = γ = 90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (V M) was 6.35 Å3 Da -1, with a solvent content of 80.62%. Initial phases were determined by molecular replacement.

AB - Human embryonically lethal abnormal vision (ELAV)-type RNA-binding protein 3 (ETR-3) has been implicated in many aspects of RNA-processing events including alternative splicing, stability, editing and translation. RNA recognition motif 3 (RRM-3) is an independent C-terminal RNA-binding domain of ETR-3 that preferentially binds to UG-rich repeats of the nuclear or cytoplasmic pre-mRNA, and along with the other domains mediates the inclusion of cardiac troponin T (c-TNT) exon 5 in embryonic muscle, which is otherwise excluded in the adult. In the present study, RRM-3 was cloned, overexpressed, purified and crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to 3 Å resolution at the home source and belonged to space group P213, with unit-cell parameters a = b = c = 118.5 Å, α = β = γ = 90°. There were two molecules of RRM-3 in the asymmetric unit and the calculated Matthews coefficient (V M) was 6.35 Å3 Da -1, with a solvent content of 80.62%. Initial phases were determined by molecular replacement.

KW - C-terminal RNA recognition motif

KW - ETR-3

KW - human ELAV-type RNA-binding protein 3

KW - RRM-3

UR - http://www.scopus.com/inward/record.url?scp=84885453821&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885453821&partnerID=8YFLogxK

U2 - 10.1107/S1744309113023439

DO - 10.1107/S1744309113023439

M3 - Article

VL - 69

SP - 1107

EP - 1109

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

SN - 1744-3091

IS - 10

ER -