| Requirements for DNA hairpin formation by RAG1/2. | |
| | |
MedLine Citation:
|
PMID: 17307873 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The rearrangement of antigen receptor genes is initiated by double-strand breaks catalyzed by the RAG1/2 complex at the junctions of recombination signal sequences and coding segments. As with some "cut-and-paste" transposases, such as Tn5 and Hermes, a DNA hairpin is formed at one end of the break via a nicked intermediate. By using abasic DNA substrates, we show that different base positions are important for the two steps of cleavage. Removal of one base in the coding flank enhances hairpin formation, bypassing a requirement for a paired complex of two signal sequences. Rescue by abasic substrates is consistent with a base-flip mechanism seen in the crystal structure of the Tn5 postcleavage complex and may mimic the DNA changes on paired complex formation. We have searched for a tryptophan residue in RAG1 that would be the functional equivalent of W298 in Tn5, which stabilizes the DNA interaction by stacking the flipped base on the indole ring. A W956A mutation in RAG1 had an inhibitory effect on both nicking and hairpin stages that could be rescued by abasic substrates. W956 is therefore a likely candidate for interacting with this base during hairpin formation. |
| | |
Authors:
|
Gabrielle J Grundy; Joanne E Hesse; Martin Gellert |
Related Documents
:
|
6548133 - An investigation into the solution structures of two self-complementary dna oligomers, ... 18201813 - Predicting ultraviolet spectrum of single stranded and double stranded deoxyribonucleic... 20932753 - Tetrasubstituted naphthalene diimide ligands with selectivity for telomeric g-quadruple... |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Intramural Date: 2007-02-16 |
Journal Detail:
|
Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 104 ISSN: 0027-8424 ISO Abbreviation: Proc. Natl. Acad. Sci. U.S.A. Publication Date: 2007 Feb |
Date Detail:
|
Created Date: 2007-03-15 Completed Date: 2007-06-13 Revised Date: 2009-11-18 |
Medline Journal Info:
|
Nlm Unique ID: 7505876 Medline TA: Proc Natl Acad Sci U S A Country: United States |
Other Details:
|
Languages: eng Pagination: 3078-83 Citation Subset: IM |
Affiliation:
|
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 5, Room 241, Bethesda, MD 20892, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
DNA
/
metabolism* DNA Breaks, Single-Stranded DNA-Binding Proteins / genetics, metabolism* Electrophoretic Mobility Shift Assay Homeodomain Proteins / genetics, metabolism* Mutagenesis Mutation / genetics Nuclear Proteins / genetics, metabolism* Nucleic Acid Conformation* |
| Chemical | |
Reg. No./Substance:
|
0/DNA-Binding Proteins; 0/Homeodomain Proteins; 0/Nuclear Proteins; 0/RAG2 protein, human; 128559-51-3/RAG-1 protein; 9007-49-2/DNA |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Genome-wide transcription map of an archaeal cell cycle.
Next Document: Night eating and obesity in the EP3R-deficient mouse.