Antibody (immunoglobulin, Ig) class switch causes B lymphocytes to switch from producing IgM to producing IgG, IgA or IgE, which improves the ability of the antibody to remove pathogens and bacterial toxins from the body. Class switching occurs by an intrachromosomal DNA recombination event that must be carefully controlled in order to avoid aberrant recombination with other chromosomes (translocations). However, occasional translocations do occur. Sometimes they result in B lymphomas or myelomas due to translocations of the Ig genes to a chromosomal site encoding an oncogene, e.g. c-myc, which induces aberrant expression of the c-myc gene. This application proposes to investigate how DNA double-strand breaks (DSBs) are introduced into the mouse c-myc gene and the role of activation-induced cytidine deaminase (AID) in the process of c-myc-IgH translocations.
In Aim 1, we will investigate the hypothesis that reactive oxygen species (ROS) induce DNA lesions that lead to DSBs in the c-myc gene, and also stimulate the deamination activity of AID, as deamination occurs by an oxidation reaction.
In Aim 2, we will investigate whether AID has additional activities besides its deaminase activity that contribute to creating c- myc-IgH translocations.
In Aim 3, we will investigate the role of the 10 amino acids at the C terminus of AID, which is required for class switch recombination but not for somatic hypermutation, and has been shown to repress c-myc-IgH translocations. We will examine the hypothesis that DNA repair proteins involved in switch recombination are recruited by the C terminus.

Public Health Relevance

This project investigates the events that initiate the generation of B cell lymphomas and myelomas. We will study how DNA breaks are introduced into the c-myc oncogene when normal mouse B cells are induced to undergo the normal process of antibody class switching in response to activation signals. These DNA breaks can result in translocations of the c-myc oncogene to the immunoglobulin (Ig) heavy (H) chain gene locus, which increases expression of the c-myc gene, and this can lead to cell transformation and malignancy. We will study whether the protein that is required for initiating DNA break formation in the IgH genes during antibody class switching (activation-induced cytidine deaminase, AID) is required for introducing DNA breaks into the c-myc gene, and whether AID has additional roles that lead to the translocations between the IgH genes and c-myc genes. We will also investigate the function of AID itself during normal class switching, as this affects translocations between the c-myc gene and the IgH locus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI088578-02
Application #
8097530
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Nasseri, M Faraz
Project Start
2010-07-01
Project End
2012-06-30
Budget Start
2011-07-01
Budget End
2012-06-30
Support Year
2
Fiscal Year
2011
Total Cost
$244,283
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Genetics
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Khair, Lyne; Guikema, Jeroen E J; Linehan, Erin K et al. (2014) ATM increases activation-induced cytidine deaminase activity at downstream S regions during class-switch recombination. J Immunol 192:4887-96
Stavnezer, Janet; Linehan, Erin K; Thompson, Mikayla R et al. (2014) Differential expression of APE1 and APE2 in germinal centers promotes error-prone repair and A:T mutations during somatic hypermutation. Proc Natl Acad Sci U S A 111:9217-22
Ucher, Anna J; Linehan, Erin K; Teebor, George W et al. (2012) The DNA glycosylases Ogg1 and Nth1 do not contribute to Ig class switching in activated mouse splenic B cells. PLoS One 7:e36061
Ranjit, Sanjay; Khair, Lyne; Linehan, Erin K et al. (2011) AID binds cooperatively with UNG and Msh2-Msh6 to Ig switch regions dependent upon the AID C terminus. J Immunol 187:2464-75
Stavnezer, Janet (2011) Complex regulation and function of activation-induced cytidine deaminase. Trends Immunol 32:194-201
Stavnezer, Janet; Bjorkman, Andrea; Du, Likun et al. (2010) Mapping of switch recombination junctions, a tool for studying DNA repair pathways during immunoglobulin class switching. Adv Immunol 108:45-109