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Diversity-generating retroelement homing regenerates target sequences for repeated rounds of codon rewriting and protein diversification.


ABSTRACT: 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 ligand-receptor interactions.

SUBMITTER: Guo H 

PROVIDER: S-EPMC2703419 | biostudies-literature | 2008 Sep

REPOSITORIES: biostudies-literature

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Diversity-generating retroelement homing regenerates target sequences for repeated rounds of codon rewriting and protein diversification.

Guo Huatao H   Tse Longping V LV   Barbalat Roman R   Sivaamnuaiphorn Sameer S   Xu Min M   Doulatov Sergei S   Miller Jeff F JF  

Molecular cell 20080901 6


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 studi  ...[more]

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