: This competitive renewal application has two related parts aimed at resolving epigenetic mechanisms and chromatin features in B cell gene control. One line of studies is directed at defining the role of novel DNA methylation at CCWGG (i.e., CmCAITGG) sites in turning off B cell gene expression in lymphocyte development and in initiating chromatin modifications (histone deacetylation and CpG methylation) in gene silencing. We recently discovered CCWGG methylation in silenced B29 genes in AIDS-related Primary Effussion Lymphoma (PEL) and myeloma cells. We also showed that CCWGG methylation blocked critical transcription factor binding and repressed transcription of the B29 promoter. We will directly test the proposition that CCWGG methylation has two related regulatory functions: 1) to turn off gene transcription, and 2) to mark genes for chromatin modifications leading to gene silencing. CCWGG methylation and chromatin modifications in gene silencing will be analyzed in B cell genes (X5, V-pre-B, mb-i) that are turned off in development. The role of CCWGG methylation in gene silencing also will be tested using chromatin integrated CCWGG methylated vs. unmethylated B29 promoter-containing transgenes. A complementary line of studies will characterize the newly identified insulator located upstream of the B29 promoter and analyze its role in controlling B cell specific B29 expression. Insulators are recently identified boundary elements that flank genes and act as barriers to external influences that could affect gene expression (i.e., silencing, activation). The B29 insulator appears to be a new type of boundary element. Its location between the B29 gene and the upstream locus control region (LCR) that regulates expression of the pituitary growth hormone gene (located downstream of 829) predicts that it shields the B29 gene from inappropriate activation/expression. We will test this predicted activity of the B29 insulator using recombinase mediated cassette exchange (RMCE) to efficiently integrate transgenes into chromatin loci. The state of the B29 insulator also will be analyzed in B29-expressing vs. silenced cells to define the role of this element in gene silencing. These studies on CCWGG methylation and insulator function address new mechanisms in B cell gene control that supercede the effects of local trans-acting factors that interact with promoters and enhancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM040185-17
Application #
6929329
Study Section
Immunobiology Study Section (IMB)
Program Officer
Marino, Pamela
Project Start
1988-05-01
Project End
2007-07-31
Budget Start
2005-08-01
Budget End
2007-07-31
Support Year
17
Fiscal Year
2005
Total Cost
$272,236
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Henson, Sarah E; Morford, Travis; Stein, Mary-Pat et al. (2011) Candidate genes contributing to the aggressive phenotype of mantle cell lymphoma. Acta Histochem 113:729-42
Kuraishy, Ali I; French, Samuel W; Sherman, Mara et al. (2007) TORC2 regulates germinal center repression of the TCL1 oncoprotein to promote B cell development and inhibit transformation. Proc Natl Acad Sci U S A 104:10175-80
Malone, Cindy S; Kuraishy, Ali I; Fike, Francesca M et al. (2006) B29 gene silencing in pituitary cells is regulated by its 3' enhancer. J Mol Biol 362:173-83
Henson, Sarah E; Tsai, Shih-Chang; Malone, Cindy Sue et al. (2006) Pir51, a Rad51-interacting protein with high expression in aggressive lymphoma, controls mitomycin C sensitivity and prevents chromosomal breaks. Mutat Res 601:113-24
Doerr, Jeanette R; Malone, Cindy S; Fike, Francesca M et al. (2005) Patterned CpG methylation of silenced B cell gene promoters in classical Hodgkin lymphoma-derived and primary effusion lymphoma cell lines. J Mol Biol 350:631-40
Patrone, Lisa; Henson, Sarah E; Wall, Randolph et al. (2004) A conserved sequence upstream of the B29 (Ig beta, CD79b) gene interacts with YY1. Mol Biol Rep 31:1-11
Gordon, Melinda S; Kanegai, Cindy M; Doerr, Jeanette R et al. (2003) Somatic hypermutation of the B cell receptor genes B29 (Igbeta, CD79b) and mb1 (Igalpha, CD79a). Proc Natl Acad Sci U S A 100:4126-31
Patrone, Lisa; Henson, Sarah E; Teodorovic, Jelena et al. (2003) Gene expression patterns in AIDS versus non-AIDS-related diffuse large B-cell lymphoma. Exp Mol Pathol 74:129-39
French, Samuel W; Malone, Cindy S; Shen, Rhine R et al. (2003) Sp1 transactivation of the TCL1 oncogene. J Biol Chem 278:948-55
Hoyer, Katrina K; French, Samuel W; Turner, Devin E et al. (2002) Dysregulated TCL1 promotes multiple classes of mature B cell lymphoma. Proc Natl Acad Sci U S A 99:14392-7

Showing the most recent 10 out of 42 publications