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Identification and solution structure of a highly conserved C-terminal domain within ORF1p required for retrotransposition of long interspersed nuclear element-1
- Source :
- The Journal of biological chemistry. 282(34)
- Publication Year :
- 2007
-
Abstract
- Long interspersed nuclear element-1 (LINE-1 or L1) retrotransposons comprise a large fraction of the human and mouse genomes. The mobility of these successful elements requires the protein encoded by open reading frame-1 (ORF1p), which binds single-stranded RNA with high affinity and functions as a nucleic acid chaperone. In this report, we have used limited proteolysis, filter binding, and NMR spectroscopy to characterize the global structure of ORF1p and the three-dimensional structure of a highly conserved RNA binding domain. ORF1p contains three structured regions, a coiled-coil domain, a middle domain of unknown function, and a C-terminal domain (CTD). We show that high affinity RNA binding by ORF1p requires the CTD and residues within an amino acid protease-sensitive segment that joins the CTD to the middle domain. Insights in the mechanism of RNA binding were obtained by determining the solution structure of the CTD, which is shown to adopt a novel fold consisting of a three-stranded beta sheet that is packed against three alpha-helices. An RNA binding surface on the CTD has been localized using chemical shift perturbation experiments and is proximal to residues previously shown to be essential for retrotransposition, RNA binding, and chaperone activity. A similar structure and mechanism of RNA binding is expected for all vertebrate long interspersed nuclear element-1 elements, since residues encoding the middle, protease-sensitive segment, and CTD are highly conserved.
- Subjects :
- Magnetic Resonance Spectroscopy
Retroelements
Molecular Sequence Data
Molecular Conformation
Retrotransposon
RNA-binding protein
Biology
Biochemistry
Protein Structure, Secondary
Mice
Open Reading Frames
Animals
Amino Acid Sequence
Molecular Biology
Genetics
Sequence Homology, Amino Acid
C-terminus
RNA
Cell Biology
Protein Structure, Tertiary
Long interspersed nuclear element
Kinetics
Long Interspersed Nucleotide Elements
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Biophysics
CTD
Domain of unknown function
Binding domain
Protein Binding
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 282
- Issue :
- 34
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....373631ccf79c405958f85baeddddb616