Journal
MOLECULAR CELL
Volume 31, Issue 6, Pages 813-823Publisher
CELL PRESS
DOI: 10.1016/j.molcel.2008.07.022
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Funding
- NIH [A1071204, A1061598]
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Diversity-generating retroelements (DGRs) introduce vast amounts of sequence diversity into target genes. During mutagenic homing, adenine residues are converted to random nucleotides in a unidirectional, reverse transcriptase-dependent transposition process from a donor template repeat (TR) to a recipient variable repeat (VR). Using a Bordetella bacteriophage DGR as a model, we demonstrate that homing occurs through a TR-containing RNA intermediate and is RecA independent. Marker transfer studies show that cDNA integration at the 3' end of VR occurs within a (G/C)14 element, and deletion analysis demonstrates that the reaction is independent of 5' end cDNA integration. cDNA integration at the 5' end of VR requires only short stretches of sequence homology. We propose that homing occurs through a unique target DNA-primed reverse transcription mechanism that precisely regenerates target sequences. This nonproliferative copy and replace mechanism enables repeated rounds of protein diversification and optimization of ligandreceptor interactions.
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