| A single amino acid substitution in ORF1 dramatically decreases L1 retrotransposition and provides insight into nucleic acid chaperone activity. | |
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MedLine Citation:
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PMID: 18790804 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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L1 is a ubiquitous interspersed repeated sequence in mammals that achieved its high copy number by autonomous retrotransposition. Individual L1 elements within a genome differ in sequence and retrotransposition activity. Retrotransposition requires two L1-encoded proteins, ORF1p and ORF2p. Chimeric elements were used to map a 15-fold difference in retrotransposition efficiency between two L1 variants from the mouse genome, T(FC) and T(Fspa), to a single amino acid substitution in ORF1p, D159H. The steady-state levels of L1 RNA and protein do not differ significantly between these two elements, yet new insertions are detected earlier and at higher frequency in T(FC), indicating that it converts expressed L1 intermediates more effectively into new insertions. The two ORF1 proteins were purified and their nucleic acid binding and chaperone activities were examined in vitro. Although the RNA and DNA oligonucleotide binding affinities of these two ORF1 proteins were largely indistinguishable, D159 was significantly more effective as a nucleic acid chaperone than H159. These findings support a requirement for ORF1p nucleic acid chaperone activity at a late step during L1 retrotransposition, extend the region of ORF1p that is known to be critical for its functional interactions with nucleic acids, and enhance understanding of nucleic acid chaperone activity. |
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Authors:
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Sandra L Martin; Diane Bushman; Fei Wang; Patrick Wai-Lun Li; Ann Walker; Jessica Cummiskey; Dan Branciforte; Mark C Williams |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. Date: 2008-09-12 |
Journal Detail:
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Title: Nucleic acids research Volume: 36 ISSN: 1362-4962 ISO Abbreviation: Nucleic Acids Res. Publication Date: 2008 Oct |
Date Detail:
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Created Date: 2008-10-14 Completed Date: 2008-11-24 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 0411011 Medline TA: Nucleic Acids Res Country: England |
Other Details:
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Languages: eng Pagination: 5845-54 Citation Subset: IM |
Affiliation:
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Department of Cell and Developmental Biology, University of Colorado School of Medicine, Aurora, CO 80045, USA. sandy.martin@ucdenver.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Substitution Animals Cell Line DNA / chemistry, metabolism DNA-Binding Proteins / genetics, metabolism* Kinetics Long Interspersed Nucleotide Elements* Mice Molecular Chaperones / genetics, metabolism Nucleic Acid Denaturation RNA / metabolism RNA-Binding Proteins / genetics, metabolism* Ribonucleoproteins / genetics, metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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GM40367/GM/NIGMS NIH HHS; GM72462/GM/NIGMS NIH HHS; P30 CA046934/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/DNA-Binding Proteins; 0/L1 ORF1 protein, human; 0/Molecular Chaperones; 0/RNA-Binding Proteins; 0/Ribonucleoproteins; 63231-63-0/RNA; 9007-49-2/DNA |
| Comments/Corrections | |
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