Abstract

Animals will be immunized with representative examples of A, B and Z-DNA and base-paired oligonucleotides of defined sequence. Resulting polyclonal and monoclonal antibodies will be tested for their ability to distinguish among: helices of different families (A, B and Z); various helical structures of one family; and various topographical features of individual helical forms. Further experiments will test the ability of the antibodies to detect unique sequence-dependent helical conformation inserted into linear or circular DNA molecules and to affect the dynamic equilibrium between different helical conformations that can be assumed by a given sequence. Antibodies will also be used to probe for corresponding unique helical regions in naturally occurring DNA, chromatin and nuclei. These experiments will provide reagents and procedures for testing the hypothesis that specific local conformational features occur within the chromatin organization of DNA and may serve as signal sites in processes regulating transcription or replication of gene information.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032375-03
Application #
3281154
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1983-07-01
Project End
1988-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Tufts University
Department
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code

  Publications

Li, Jing; Geissal, Erik D; Li, Wenqin et al. (2005) Repertoire diversification in mice with an IgH-locus-targeted transgene for the rearranged VH domain of a physiologically selected anti-ssDNA antibody. Mol Immunol 42:1475-84
Jang, Y J; Stollar, B D (2003) Anti-DNA antibodies: aspects of structure and pathogenicity. Cell Mol Life Sci 60:309-20
O'Connor, K C; Nguyen, K; Stollar, B D (2001) Recognition of DNA by VH and Fv domains of an IgG anti-poly(dC) antibody with a singly mutated VH domain. J Mol Recognit 14:18-28
Li, J; Fernandez, L; O'Connor, K C et al. (2001) The rearranged V(H) domain of a physiologically selected anti-single-stranded DNA antibody as a precursor for formation of IgM and IgG antibodies to diverse antigens. J Immunol 167:3746-55
O'Connor, K C; Ghatak, S; Stollar, B D (2000) Use of hydrophobic interaction chromatography to separate recombinant antibody fragments from associated bacterial chaperone protein GroEL. Anal Biochem 278:239-41
Stollar, B D (2000) Contributions of antibody VH domains to anti-DNA autoreactivity. Clin Rev Allergy Immunol 18:41-50
Wang, X; Stollar, B D (2000) Human immunoglobulin variable region gene analysis by single cell RT-PCR. J Immunol Methods 244:217-25
Chen, Y; Stollar, B D (1999) DNA binding by the VH domain of anti-Z-DNA antibody and its modulation by association of the VL domain. J Immunol 162:4663-70
Lecerf, J M; Chen, Y; Richalet-Secordel, P et al. (1998) Autoreactivity of human VH domains from cDNA libraries: analysis with a bacterial expression system. J Immunol 161:1274-83
Jang, Y J; Sanford, D; Chung, H Y et al. (1998) The structural basis for DNA binding by an anti-DNA autoantibody. Mol Immunol 35:1207-17

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