A great deal has been learned about the biochemistry of activation induced cytidine deaminase (AID) and the process of somatic hypermutation of antibody variable regions, but the rules that govern the preferential targeting of AID to hot spots within the antibody V regions and the mechanisms that are responsible that targeting are still obscure. Here we propose to use the Ramos Burkitt's lymphoma cell line to examine the role of local DNA sequences and structures in recruiting AID to the variable region. We will also examine whether particular chromatin modifications play a role in targeting AID to different parts of the V region exon and its immediate flanking sequences and compare them to the switch regions. We will use recombinant mediated cassette exchange to manipulated the structure of the variable region in the endogenous heavy chain locus and chromatin immunoprecipitation to identify the chromatin modifications and the sites of interaction of AID with the heavy chain variable region. We will use purified AID and computational tools that we have developed to compare the preferred sites of action of AID in vitro and to those that are targeted in Ramos cells and to a large data base of human V region mutations that have arisen in vivo.

Public Health Relevance

In order to protect ourselves from pathogenic organisms and their products, we produce a wide variety of antibodies that become more effective as the immune response progresses. This affinity maturation of the immune response and the ability of the antibodies to carry out many different functions throughout the body depends a highly mutagenic process that occurs in the B cells making the antibody and is largely restricted to the antibody genes. Here we will examine how this process is selectively targeted to the antibody genes rather than mistargeted to other genes where it would lead to the malignant transformation of the antibody forming cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA072649-15
Application #
8465102
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Mccarthy, Susan A
Project Start
1997-09-15
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
15
Fiscal Year
2013
Total Cost
$371,121
Indirect Cost
$147,554
Name
Albert Einstein College of Medicine
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Wang, Xiaohua; Fan, Manxia; Kalis, Susan et al. (2014) A source of the single-stranded DNA substrate for activation-induced deaminase during somatic hypermutation. Nat Commun 5:4137
Jaszczur, Malgorzata; Bertram, Jeffrey G; Pham, Phuong et al. (2013) AID and Apobec3G haphazard deamination and mutational diversity. Cell Mol Life Sci 70:3089-108
Schaetzlein, Sonja; Chahwan, Richard; Avdievich, Elena et al. (2013) Mammalian Exo1 encodes both structural and catalytic functions that play distinct roles in essential biological processes. Proc Natl Acad Sci U S A 110:E2470-9
Chahwan, Richard; van Oers, Johanna M M; Avdievich, Elena et al. (2012) The ATPase activity of MLH1 is required to orchestrate DNA double-strand breaks and end processing during class switch recombination. J Exp Med 209:671-8
Chahwan, Richard; Edelmann, Winfried; Scharff, Matthew D et al. (2011) Mismatch-mediated error prone repair at the immunoglobulin genes. Biomed Pharmacother 65:529-36
Chahwan, Richard; Wontakal, Sandeep N; Roa, Sergio (2011) The multidimensional nature of epigenetic information and its role in disease. Discov Med 11:233-43
Baughn, Linda B; Kalis, Susan L; MacCarthy, Thomas et al. (2011) Recombinase-mediated cassette exchange as a novel method to study somatic hypermutation in Ramos cells. MBio 2:
Peled, Jonathan U; Sellers, Rani S; Iglesias-Ussel, Maria D et al. (2010) Msh6 protects mature B cells from lymphoma by preserving genomic stability. Am J Pathol 177:2597-608
Ramachandran, Shaliny; Chahwan, Richard; Nepal, Rajeev M et al. (2010) The RNF8/RNF168 ubiquitin ligase cascade facilitates class switch recombination. Proc Natl Acad Sci U S A 107:809-14
van Oers, Johanna M M; Roa, Sergio; Werling, Uwe et al. (2010) PMS2 endonuclease activity has distinct biological functions and is essential for genome maintenance. Proc Natl Acad Sci U S A 107:13384-9

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