Activation-induced cytidine deaminase deaminates deoxycytidine on single-stranded DNA but requires the action of RNase

Ronda Bransteitter, Phuong Pham, Matthew D. Scharff, Myron F. Goodman

Research output: Contribution to journalArticle

516 Citations (Scopus)

Abstract

The expression of activation-induced cytidine deaminase (AID) is prerequisite to a "trifecta" of key molecular events in B cells: class-switch recombination and somatic hypermutation in humans and mice and gene conversion in chickens. Although this critically important enzyme shares common sequence motifs with apolipoprotein B mRNA-editing enzyme, and exhibits deaminase activity on free deoxycytidine in solution, it has not been shown to act on either RNA or DNA. Recent mutagenesis data in Escherichia coli suggest that AID may deaminate dC on DNA, but its putative biochemical activities on either DNA or RNA remained a mystery. Here, we show that AID catalyzes deamination of dC residues on single-stranded DNA in vitro but not on double-stranded DNA, RNA-DNA hybrids, or RNA. Remarkably, it has no measurable deaminase activity on single-stranded DNA unless pretreated with RNase to remove inhibitory RNA bound to AID. AID catalyzes dC → dU deamination activity most avidly on double-stranded DNA substrates containing a small "transcription-like" single-stranded DNA bubble, suggesting a targeting mechanism for this enigmatic enzyme during somatic hypermutation.

Original languageEnglish (US)
Pages (from-to)4102-4107
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume100
Issue number7
DOIs
StatePublished - Apr 1 2003

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Deoxycytidine
Single-Stranded DNA
Ribonucleases
DNA
RNA
Deamination
Gene Conversion
Double-Stranded RNA
Enzymes
Mutagenesis
Genetic Recombination
AICDA (activation-induced cytidine deaminase)
Chickens
B-Lymphocytes
Escherichia coli

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Activation-induced cytidine deaminase deaminates deoxycytidine on single-stranded DNA but requires the action of RNase. / Bransteitter, Ronda; Pham, Phuong; Scharff, Matthew D.; Goodman, Myron F.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, No. 7, 01.04.2003, p. 4102-4107.

Research output: Contribution to journalArticle

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