DNA Repair Labortoary
Lab head: Dr Chris Jolly
Location: Centenary Institute
My group studies antibody mutation in activated B cells, which is initiated by the DNA editing enzyme "AID". B cells mutate their antibody genes at extremely high rates during infections, to rapidly optimise the ability of the antibodies they make to neutralise the infecting pathogen. "Off-target" many adult B cell cancers, so we seek to understand why AID-induced DNA damage leads to mutation, when similar DNA damage is generally repaired faithfully.
Funding: NHMRC, Cancer Council NSW
Research approach equipment: We use retroviral-mediated transgenesis of mouse haematopoietic cells to modify DNA repair pathways in B cells, in order to determine how inactivating or cell cycle restricting DNA repair pathways influences gene mutation in activated B cells.
Chan, T.D., K. Wood, J.R. Hermes, D. Butt, C.J. Jolly, A. Basten, and R. Brink. 2012. Elimination of germinal center-derived self-reactive B cells is governed by the location and level of self-antigen. Immunity in press.
Sharbeen, G., C.W. Yee, A.L. Smith, and C.J. Jolly. 2012. Ectopic restriction of DNA repair reveals that UNG2 excises AID-induced uracils predominantly or exclusively during G1 phase. J Exp Med 209:965-974. http://www.ncbi.nlm.nih.gov/pubmed/22529268
Sharbeen, G., A.J. Cook, K.K. Lau, J. Raftery, C.W. Yee, and C.J. Jolly. 2010. Incorporation of dUTP does not mediate mutation of A:T base pairs in Ig genes in vivo. Nucleic Acids Res. 38:8120-8130. http://www.ncbi.nlm.nih.gov/pubmed/20705648
Jolly, C.J., A.J. Cook, and J.P. Manis. 2008. Fixing DNA breaks during class switch recombination. J Exp Med. 205:509-513. http://www.ncbi.nlm.nih.gov/pubmed/18332183
Cook, A.J., J.M. Raftery, K.K. Lau, A. Jessup, R.S. Harris, S. Takeda, and C.J. Jolly. 2007. DNA-Dependent Protein Kinase Inhibits AID-Induced Antibody Gene Conversion. PLoS Biol. 5:792-799. http://www.ncbi.nlm.nih.gov/pubmed/17355182
Jolly, C., A.J. Cook, J. Raferty, and M.E. Jones. 2007. Measuring bidirectional mutation. J. Theor. Biol. 246:269-277. http://www.ncbi.nlm.nih.gov/pubmed/17292922
Cook, A.J.L., L. Oganesian, P. Harumal, A. Basten, R. Brink, and C.J. Jolly. 2003. Reduced Switching in SCID B Cells Is Associated with Altered Somatic Mutation of Recombined S Regions. J Immunol 171:6556-6564. http://www.ncbi.nlm.nih.gov/pubmed/14662857
Jolly, C.J., and M.S. Neuberger. 2001. Somatic hypermutation of immunoglobulin kappa transgenes: association of mutability with demethylation. Immunol. Cell Biol. 79:18-22. http://www.ncbi.nlm.nih.gov/pubmed/21100930
Williams, G.T., C.J. Jolly, J. Kohler, and M.S. Neuberger. 2000. The contribution of somatic hypermutation to the diversity of serum immunoglobulin: dramatic increase with age. Immunity 13:409-417. http://www.ncbi.nlm.nih.gov/pubmed/11021538
Jolly, C.J., S.D. Wagner, C. Rada, N. Klix, C. Milstein, and M.S. Neuberger. 1996. The targeting of somatic hypermutation. Semin. Immunol. 8:159-168. http://www.ncbi.nlm.nih.gov/pubmed/8738915
Klix, N., C.J. Jolly, S.L. Davies, M. Bruggemann, G.T. Williams, and M.S. Neuberger. 1998. Multiple sequences from downstream of the J kappa cluster can combine to recruit somatic hypermutation to a heterologous, upstream mutation domain. Eur. J. Immunol. 28:317-326. http://www.ncbi.nlm.nih.gov/pubmed/9485211
Neuberger, M.S., M.R. Ehrenstein, N. Klix, C.J. Jolly, J. Yelamos, C. Rada, and C. Milstein. 1998. Monitoring and interpreting the intrinsic features of somatic hypermutation. Immunol. Rev. 162:107-116. http://www.ncbi.nlm.nih.gov/pubmed/9602357
Screening for drugs to inhibit AID, a key contributor to B cell neoplasia
Primary supervisor: Christopher Jolly
During responses to infection or immunisation, antibody-producing “B” cells mutate their antibody genes at extreme rates. Rare mutations that improve antibodies are selected by competition between B cells favouring those which make the better antibodies: Darwinian evolution on extreme “fast-forward”. We aim to understand this process because it is essential for normal immunity and effective vaccination, but also because when it goes wrong, it causes aggressive human cancers such as Burkitt’s lymphoma and Multiple myeloma.
Both physiological and oncogenic gene mutation in B cells is induced by the DNA editing enzyme "AID". The aim of this project is to develop an assay to identify drugs with potential to inhibit AID. Two approaches will be tested: (1) a test-tube assay that directly measures AID activity in a high-thoughput cell-free system, and (2) a lower throughput cell-based assay that indirectly measures AID activity and detects cytotoxic drug affects at the same time.
The project will involve the production of recombinant AID in bacterial cells, optimisation of high-throughput 96-well scintillation assays, and high-throughput flow cytometry.
Discipline: Infectious diseases and Immunology
Keywords: B cells, Cancer, Cell & Molecular Biology
Contact: Email Christopher Jolly