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Proc Natl Acad Sci U S A. 2013 May 14;110(20):8146-51. doi: 10.1073/pnas.1303126110. Epub 2013 Apr 29.

Mechanism of somatic hypermutation at the WA motif by human DNA polymerase η.

Author information

1
Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China.

Abstract

Somatic hypermutation is programmed base substitutions in the variable regions of Ig genes for high-affinity antibody generation. Two motifs, RGYW and WA (R, purine; Y, pyrimidine; W, A or T), have been found to be somatic hypermutation hotspots. Overwhelming evidence suggests that DNA polymerase η (Pol η) is responsible for converting the WA motif to WG by misincorporating dGTP opposite the templating T. To elucidate the molecular mechanism, crystal structures and kinetics of human Pol η substituting dGTP for dATP in four sequence contexts, TA, AA, GA, and CA, have been determined and compared. The T:dGTP wobble base pair is stabilized by Gln-38 and Arg-61, two uniquely conserved residues among Pol η. Weak base paring of the W (T:A or A:T) at the primer end and their distinct interactions with Pol η lead to misincorporation of G in the WA motif. Between two WA motifs, our kinetic and structural data indicate that A-to-G mutation occurs more readily in the TA context than AA. Finally, Pol η can extend the T:G mispair efficiently to complete the mutagenesis.

KEYWORDS:

A-to-G transition; immunoglobulin; π–cation stacking

PMID:
23630267
PMCID:
PMC3657764
DOI:
10.1073/pnas.1303126110
[Indexed for MEDLINE]
Free PMC Article
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