Identification of gemin5 as a novel 7-methylguanosine cap-binding protein

Shelton Bradrick, Matthias Gromeier

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Background: A unique attribute of RNA molecules synthesized by RNA polymerase II is the presence of a 7-methylguanosine (m7G) cap structure added co-transcriptionally to the 5′ end. Through its association with trans-acting effector proteins, the m7G cap participates in multiple aspects of RNA metabolism including localization, translation and decay. However, at present relatively few eukaryotic proteins have been identified as factors capable of direct association with m7G. Methodology/Principal Findings: Employing an unbiased proteomic approach, we identified gemin5, a component of the survival of motor neuron (SMN) complex, as a factor capable of direct and specific interaction with the m7G cap. Gemin5 was readily purified by cap-affinity chromatography in contrast to other SMN complex proteins. Investigating the underlying basis for this observation, we found that purified gemin5 associates with m7G-linked sepharose in the absence of detectable eIF4E, and specifically crosslinks to radiolabeled cap structure after UV irradiation. Deletion analysis revealed that an intact set of WD repeat domains located in the N-terminal half of gemin5 are required for cap-binding. Moreover, using structural modeling and site-directed mutagenesis, we identified two proximal aromatic residues located within the WD repeat region that significantly impact m7G association. Conclusions/Significance: This study rigorously identifies gemin5 as a novel cap-binding protein and describes an unprecedented role for WD repeat domains in m7G recognition. The findings presented here will facilitate understanding of gemin5's role in the metabolism of non-coding snRNAs and perhaps other RNA pol II transcripts.

Original languageEnglish (US)
Article numbere7030
JournalPLoS One
Volume4
Issue number9
DOIs
StatePublished - Sep 14 2009
Externally publishedYes

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RNA Cap-Binding Proteins
SMN Complex Proteins
binding proteins
RNA Polymerase II
Association reactions
RNA
motor neurons
Metabolism
Affinity chromatography
Mutagenesis
metabolism
proteins
site-directed mutagenesis
DNA-directed RNA polymerase
affinity chromatography
Site-Directed Mutagenesis
Affinity Chromatography
Proteomics
proteomics
Sepharose

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Identification of gemin5 as a novel 7-methylguanosine cap-binding protein. / Bradrick, Shelton; Gromeier, Matthias.

In: PLoS One, Vol. 4, No. 9, e7030, 14.09.2009.

Research output: Contribution to journalArticle

Bradrick, Shelton ; Gromeier, Matthias. / Identification of gemin5 as a novel 7-methylguanosine cap-binding protein. In: PLoS One. 2009 ; Vol. 4, No. 9.
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