Mutational activity in cell line WEHI-231.
The cell line WEHI-231 expresses activation-induced cytidine deaminase (AID), the enzyme that mediates hypermutation and immunoglobulin class switch recombination in activated B cells. Although both the cDNA sequence and protein expression of AID appear normal, the frequency of mutation at the endogenous immunoglobulin locus is low. In this report, we have tested the mutational activity of the cell line with three different indicator constructs. The first construct measures a composite rate of transversions of C to G and C to A, respectively. The second construct measures only transversion from C to G. The third measures the canonical AID activity, from C to U, which after cell replication can result in a C to T transition. We found that in WEHI-231, the C to G activity is 32- to 37-times lower than in the hypermutating cell line 18-81. The C to T activity is also much reduced, but only 12-fold. We suggest that the WEHI-231 lacks an activity that subverts the faithful repair of incipient C to U mutations.
Wang
Clifford L
CL
Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA. mutator@itsa.ucsf.edu
Wabl
Matthias
M
D000818:Animals; D002460:Cell Line; D003564:Cytidine Deaminase / Q000378:metabolism*; D017930:Genes, Reporter; D006801:Humans; D009154:Mutation; D013997:Time Factors; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase